JUNE / JULY 2021 # 98

Upfront In My View Feature Sitting Down With Algal doom and Alzheimer’s Sample preparation takes Citizen science: a new era in Food safety fanatic, disease inclusion center stage analytical chemistry? Michel Nielen

06 12 26 – 37 50 – 51

The (Long and Winding) Road to LPGC-MS

Three trailblazers recall the arduous journey to low-pressure GC 16 – 25

www.theanalyticalscientist.com Shimadzu_TAS_062021.qxp_210x266 19.04.21 12:14 Seite 1

Suits perfectly Grows according to needs

The brand-new AOC-30 series of automatic sample injection Grows flexibly with lab’s future needs systems for gas chromatographs benefits routine analysis work through customizable accessory set-up options and many in pharmaceutical, chemical and environmental. Available as upgrade possibilities Single and Dual Tower solution, the AOC-30 series achieves high throughput with a capacity of up to 150 vials. Equipped Expert-level measurement results with unique technologies and functionalities, the automatic ensured by “Analytical Intelligence” functionality started with sample injection system is suited perfectly to automation, just a click productivity and remote operability needs of next-generation laboratories. Application-specific configuration through various syringe sizes allowing flexible injection volumes Precise results and long-term operation based on features such as intelligent syringe wash and Xtra live septum for extended service life

www.shimadzu.eu/suits-perfectly The Analytical Intelligence logo is a trademark of Shimadzu Corporation. Shimadzu_TAS_062021.qxp_210x266 19.04.21 12:14 Seite 1

A Democracy for Discovery Editorial Could citizen-driven analytical science supercharge change?

t’s not hard to see why science appears somewhat elitist to outsiders. Those among its upper echelons typically hold multiple degrees in challenging subjects from prestigious Iinstitutions, often with awards lists as long as your arm. And that’s not a criticism, mind you – just an observation. Nevertheless, I can’t help but wonder if this status quo underscores much of the disconnect we see between scientists and the public... One trend that can bridge these two groups? Citizen science. Described as “the democratization of science” by Michel Nielen in this month’s second feature (page 32), it could help bring research access to the masses. Today’s public is welcome (and even encouraged) to contribute to studies on all manner of subjects – from monitoring the effect of light pollution on the visibility of constellations in the night sky to a good old-fashioned avian census. Analytical science, by its very nature, must take the idea one step further; for example, by empowering the citizens with smartphone-linked mini-machines that can assess food safety or detect disease. But who are the winners in this relationship? What we’re talking about is a symbiosis; the general public buys into and directly benefits from the hard work of scientists, meanwhile the scientists (hopefully) benefit from previously unobtainable pools of data and renewed (or newfound!) interest in the research subjects they hold so dear. In the age of anti-maskers and climate change-deniers, perhaps such mutually beneficial interaction could exert effects that reach far beyond what one might first anticipate. After all, we must all work together if we are to tackle the great crises facing humanity. Green chemistry is almost certain to play a pivotal role in our field’s own contributions to sustainability – see what Elena Ibañez of the Sample Preparation Study Group and Network has to say about that on page 16. Suits perfectly Innovation will also be needed to guide us to a brighter (or simply tolerable) tomorrow – the need for measurement science Grows according to needs can only significantly and continuously increase. Could low- The brand-new AOC-30 series of automatic sample injection Grows flexibly with lab’s future needs pressure GC give us the speed and simplicity we need to ramp systems for gas chromatographs benefits routine analysis work through customizable accessory set-up options and many up testing efforts to unprecedented levels? Perhaps – but turn in pharmaceutical, chemical and environmental. Available as upgrade possibilities to page 20 to make up your own mind. Single and Dual Tower solution, the AOC-30 series achieves I, for one, am excited to see how analytical – and citizen – high throughput with a capacity of up to 150 vials. Equipped Expert-level measurement results science evolve in the future. After all, working at The Analytical with unique technologies and functionalities, the automatic ensured by “Analytical Intelligence” functionality started with sample injection system is suited perfectly to automation, just a click Scientist certainly grants us wonderful insights – but not the productivity and remote operability needs of next-generation power or tools to wield analytical science for personal ends! laboratories. Application-specific configuration (Could somebody please let me know when smartphone through various syringe sizes allowing flexible injection volumes spectrometers are about to hit the consumer market?) Precise results and long-term operation based on features such as intelligent syringe wash and Xtra live septum for extended service life Matthew Hallam Editor

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In My View 08 12 Giorgia Purcaro discusses the importance of sample prep in multidimensional analysis

03 Editorial Upfront 14 There are a number of key A Democracy for Discovery, considerations chemists must by Matthew Hallam 06 Quick reads on the latest take into account to optimize research, from an answer to sample prep success - Verónica the mysterious eagle deaths Pino shares why material choice On The Cover plaguing the US, to a cheap and is one of them rapid test for COVID-19 Despite the obstacles, 16 Elena Ibañez discusses the LPGC has persevered. importance of green analytical Here, we chose to portray this chemistry in meeting the (arduous) journey with a classic Sustainable Development Goals image of endurance. set out by the UN ISSUE 98 - JUNE / JULY 2021

Feel free to contact any one of us: [email protected] Content Team Editor - Matthew Hallam Lauren Robertson (Deputy Editor) Frank van Geel (Scientific Director) Commercial Team Publisher - Lee Noyes Gaurav Avasthi (Business Development Manager) Nick Ransom (Business Development Executive Americas) Design Team Head of Design - Marc Bird Hannah Ennis (Senior Designer) Charlotte Brittain (Designer)

Digital Team Digital Team Lead - David Roberts Peter Bartley (Digital Producer Web/Email) Abygail Bradley (Digital Producer Web/App) Audience Team Audience Growth Strategy Manager – Brice Agamemnon CRM & Compliance CRM & Compliance Manager - Tracey Nicholls Hayley Atiz (CRM Assistant) Commercial Support Team Internal Systems Manager - Jody Fryett Dan Marr (Campaign Reporting Analyst) Commercial Services 32 Commercial Service and Social Media Manager - Matt Everett Kevin O'Donnell (Marketing Executive) Alice Daniels-Wright (Video Project Manager) Jess Lines (Project Support Coordinator) Lindsey Vickers (Sales Support Project Manager) Jennifer Bradley (Sales Support Coordinator) Marketing Team Features Departments Marketing Manager - Katy Pearson Jo Baylay (Marketing Executive) 20 The (Long and Winding) Road 42 Solutions: NIST research Accounts Team Kerri Benson (Accounts Assistant) to LPGC-MS chemist Melissa Phillips tells us Emily Scragg (Accounts Apprentice)

Three trailblazers on LPGC’s about the aims of their cannabis Human Resources journey recount the obstacles quality assurance program Human Resource Manager - Tara Higby that sprung up during Management Team Chief Executive Officer - Andy Davies development, and share how the Chief Operating Officer - Tracey Peers Senior Vice President (North America) - Fedra Pavlou technique managed to persevere Financial Director - Phil Dale Sitting Down With Commercial Director - Richard Hodson Content Director - Rich Whitworth 32 Empowering the Analytical Scientist in Everyone 50 Michel Nielen, Principal Change of address [email protected] Michel Nielen wants to bring Scientist of Wageningen Food Hayley Atiz, The Analytical Scientist, Texere Publishing Limited, science to the masses. The Safety Research (WFSR) Booths Park 1, Chelford Road, Knutsford, Cheshire, WA16 8GS, UK General enquiries advantages are clear, but what and Professor of Analytical www.texerepublishing.com | [email protected] obstacles do we face? Chemistry at Wageningen +44 (0) 1565 745 200 | [email protected] Distribution: TheAnalytical Scientist (ISSN 2051-4077), University, Wageningen, the is published monthly by Texere Publishing Limited, Booths Park 1, Chelford Road, Knutsford, Cheshire, WA16 8GS, UK. Netherlands Single copy sales $15 (plus postage, cost available on request [email protected]). Non-qualified annual subscription cost is available on request.

Reprints & Permissions – [email protected] The copyright in the materials contained in this publication and the typographical arrangement of this publication belongs to Texere Publishing Limited. No person may Editorial California at Berkeley, USA Biosystems Research Peter Schoenmakers, copy, modify, transmit, distribute, display, reproduce, publish, licence or create works from Gary Hieftje, Indiana Institute, USA University of Amsterdam, any part of this material or typographical arrangement, or otherwise use it, for any public Advisory or commercial use without the prior written consent of Texere Publishing Limited. Board University, USA (Retired) Luigi Mondello, University The Netherlands The names, publication titles, logos, images and presentation style appearing in this Hans-Gerd Janssen, of Messina, Italy Robert Kennedy, publication which identify Texere Publishing Limited and/or its products and services, including but without limitation Texere and The Analytical Scientist are proprietary Chris Harrison, San Diego Unilever Research and Martin Gilar, Waters, USA University of Michigan, marks of Texere Publishing Limited. Nothing contained in this publication shall be State University, USA Development, The Michelle Reid, Cristal USA deemed to confer on any person any licence or right on the part of Texere Publishing Christina Jones, Research Netherlands Therapeutics, The Ron Heeren, Maastricht Limited with respect to any such name, title, logo, image or style. chemist, NIST, USA Ian Wilson, Imperial Netherlands University, The The Analytical Scientist is printed using soy ink Emily Hilder, University of College London, UK Monika Dittmann, Netherlands South Australia, Australia Jenny Van Eyk, Director Independent Analytical Samuel Kounaves, Tufts Frantisec Svec, University of of the Advanced Clinical Scientist, Germany University, USA 6 Upfront Upfront

Research Algal Doom Innovation Trends After 25 years, we might finally have an explanation for the mysterious deaths of bald eagles across America

When bald eagles started dropping dead in Arkansas in the mid-90s, scientists were stumped – in part because of the strange ways that these birds acted. Some flew into cliff faces. Others stumbled about drunkenly on the ground. “These affected waters had one thing MS imaging was used to analyze the Suspected causes from known toxins to in common: dense aquatic vegetation, cyanobacterium in its true environment. infectious disease were explored, but a especially Hydrilla.” But there were To the researchers’ delight, a metabolite definitive answer remained elusive. areas where Hydrilla grew and no AVM absent from the laboratory cultures Twenty-five years later, we may finally occurred, so what factor made the deadly was detected. “The metabolite was have one… difference in some places? Upon closer spectacular,” Timo explains. “The sum The first clue? A cause of death: avian inspection, a previously unidentified formula was unknown, but it contained vacuolar myelinopathy (AVM). The birds cyanobacterium was found on the five bromine atoms. This explained why were dying because of lesions in their Hydrilla in affected lakes. A suspect at the metabolite wasn’t produced in the lab brains and spinal cords. “We know of long last… – there was no bromine available there!” more than 130 bald eagles that have died “I received a sample of the cyanobacterium Analysis using NMR, infrared, high- since the discovery of AVM in 1994,” says in 2010,” Timo told us. “We purified resolution MS, and X-ray crystallography Timo Niedermeyer, a researcher from it, but (extremely) slow growth of the then dove a little deeper. Germany’s Martin Luther University strain meant that it took us 18 months to From there, another team was able to who tackled the bizarre events. “When accumulate enough of it to feed to chickens show that this novel toxin caused AVM the disease first arose, it killed around and test our theory.” After all that time, with relative ease! But that’s not the end 70 percent of the bald eagles wintering the cyanobacteria did not cause AVM in of this story. “Now we need to find out at DeGray Lake, and it was later found the chickens and the project was almost where this bromide is coming from,” Timo to affect waterfowl, fish, amphibians, discontinued. But then Timo’s team had explains. “Higher bromide concentrations reptiles, and crustaceans, too!” another thought: what if the cyanobacteria are unusual in freshwater and, though it Therein lies clue number two: proximity present in the lakes produced different may have natural origins, it’s likely that to specific bodies of water. Timo says, metabolites to those grown in the lab? human activities are to blame.”

INFOGRAPHIC

WHAT IS FOOD FRAUD? MOST COMMONLY There are a number of definitions out ADULTERATED Fare Share there, but the European Commission FOODS defines food fraud as“any suspected intentional action by businesses We explore the stats or individuals for the purpose of Fish/ seafood behind food fraud deceiving purchasers and gaining Dairy across the globe undue advantage.” It encompasses Meat adulteration, tampering, product overrun, theft, diversion, simulation Alcoholic beverages and counterfeiting. Oils/fats BUSINESS IN BRIEF More Secrets of • Jim Waters, founder and leader of Waters Corporation, sadly the Mary Rose passed on May 17, 2021 at 95 years of age. His invaluable Henry VIII’s prized flagship contributions to the analytical had a multi-ethnic crew, universe are evident in the feats according to multi-isotope achieved in labs around the analysis of teeth world each and every day (1). • Thermo Fisher has joined forces dimensional molecular Multi-isotope analysis of remains from with the UK’s University of structures. The product will the Mary Rose has offered insight into Sheffield to develop end-to- likely find a home in drug the true diversity of Tudor England (1); end workflows to characterize discovery, synthetic chemistry, specifically, the researchers combined and monitor oligonucleotide and material science (4). strontium (87Sr/86Sr), oxygen (δ18O), and mRNA products. The • Shimadzu and Shionogi have sulfur (δ34S), carbon (δ13C) and approaches will combine signed a basic agreement to nitrogen (δ15N) isotope analysis of magnetic bead sample work towards a wastewater- dental samples to reconstruct childhood preparation technology with based early detection system diet and origins of eight crew members. high-resolution accurate MS to for the monitoring of infectious There has been little isotope analysis assess emerging vaccines and diseases, including SARS- performed on post-medieval remains drug products (2). CoV-2. Increased sensitivity in Britain, so it is interesting that the • Peak Scientific have partnered will be key to the success provenance isotope data presented with eForests, a UK-based of these methods in eastern suggests that up to three crew members not-for-profit, with the aim countries like Japan, where were from warmer climes than Britain. of planting a tree for each gas cases are much lower than the Co-author Alexzandra Hildred, said generator sold as of April 2021. US and Europe; the partnership (2): “We never expected the diversity The initiative supports the will tackle such challenges head to be so rich. This study transforms our company’s mission to improve on (5). perceived ideas regarding the composition sustainability across their of the nascent English navy.” operations (3). References • Jeol and Rigaku have launched 1. https://bit.ly/3gC9H5e References the XtaLAB Synergy-ED 2. https://bit.ly/35y366F 1. J Scorrer et al., R Soc Open Sci, 8 (2021). – a new and fully integrated 3. https://bit.ly/3gwTrDC DOI : 10.1098/rsos.202106 electron diffractometer for 4. https://bit.ly/3iSq4x5 2. The Mary Rose (2021). Available at : the determination of three- 5. https://bit.ly/3q2tajG https://maryrose.org/news/multicultural/

MOST NOTORIOUS FOOD FRAUD CASES

1981 Olive oil 2007 Pet food 2008 Melamine milk 2009 Peanuts 2013 Horsemeat Industrial rapeseed Melamine would make A number of Chinese Seven people died and Mostly affecting the oil was contaminated a bigger splash in a companies were found hundreds fell ill when UK and Ireland, the with the toxic year’s time, but it first to be watering down the Peanut Corporation “horsemeat scandal” compound, aniline, hit the papers after their milk and adding of America knowingly arose when equine and sold as “olive oil” thousands of pets in melamine to bypass shipped salmonella- DNA was found in to unwitting street the US died because protein tests. This led contaminated peanut frozen beefburgers vendors across Spain. they’d been given to over 50,000 children butter across the US. sold in several This led to more than food tainted with the being hospitalized. supermarkets. 1000 deaths. compound.

www.theanalyticalscientist.com 8 Upfront

Alzheimer’s Disease: Inclusion Matters

Renã Robinson tells us about her ongoing research into the importance of diverse patient groups in Alzheimer’s studies

Can you tell me a bit about your recent significant differences in the proteome What’s the biggest takeaway from work in Alzheimer’s disease? between people from different ethnic this research? Over the last few months we’ve really groups, but it also highlighted that The biggest thing we’ve learned from these been advancing our efforts to use a majority of pathways seem pretty studies is that you absolutely have to look at proteomics and lipidomics for testing consistent regardless of who presents a lot of people across diverse ethnic groups. biospecimens from representative with Alzheimer’s disease. A lot of what we know about disease has samples of underrepresented groups – come from a European-centric, non- mostly African American and Black What are the next steps for your research? Hispanic white cohort. Before we can even adults. Our recent work kicked off We are now equipping ourselves to generalize within the African American with a pilot paper we published that handle larger numbers of samples from community, we need a broader set of looks at postmortem brain tissue from studies with a significant representation individuals to partake in these studies a small cohort of African Americans of African American, Hispanic and to ensure they are as representative as and non-Hispanic whites. The aim was Latinx adults. The core driver behind possible. This doesn’t just benefit African to determine whether we could see any this is an NIH grant that was funded Americans; it benefits us all by ensuring we differences in protein biomarkers when to enable us to study hypertension as a better understand the disease as a whole. we stratified based on how individuals risk factor for AD and to examine how self-reported their race. This gave us the biochemical markers can be used to Read the full interview online: initial insight that there might be some evaluate that risk. https://bit.ly/35HEPLw

researchers tried to rectify this by the brand of crayon. Overall, the synthetic Measuring performing a multi-analytical study and organic pigments used in both brands of 44 crayons belonging to revered were largely similar, with the exception of Munch’s Crayons expressionist artist Edvard Munch. the greens; these were made using either The researchers used a combination Prussian blue and chrome yellow, or a mix Analysis of the artist’s pastel of X-ray fluorescence spectrometry, FT- of viridian, ultramarine and lead chromate. crayons provides a rare glimpse IR, Raman, surface-enhanced Raman The researchers believe the detailed into a favored tool that is often spectroscopy, pyrolysis GC-MS, LC- results will aid conservation strategies overlooked in technical studies MS and HPLC-HRMS to elucidate the for both Munch’s and other 19th century composition of the crayons. artists’ works. Unlike their counterparts, pastel The results showed the crayons were crayons appear to have been neglected oil-based, with the main binding media Reference in technical studies into the composition being beeswax, palm oil or Japan wax in 1. J La Nasa et al., Scientific Reports, 11 (2021). of modern painting materials. Recently, different relative abundances dependent on DOI : 10.1038/s41598-021-86031-6 Upfront 9

IMAGE OF THE MONTH

Dime-a-Dozen Diagnosis

A low-cost spectroscopy- based diagnostic could make fast (and accurate) COVID-19 testing more accessible

Current methods for SARS-CoV-2 detection rely heavily on RT-PCR – an admittedly highly accurate but Silent Spread: COVID-19 in Art costly technique that requires trained personnel and significant lab space – For the past year, artist Rebecca Kamen has worked with microscopists from the and it cannot be described as rapid. National Institutes of Health and other researchers to create a series of artwork that Now, researchers at Penn Medicine explores the concept of “corona” at both the micro and macro levels. The image above have developed a new smartphone- is a sneak preview of one such sculpture, but her new exhibition will also premiere a based test for COVID-19 that delivers large wall-mounted installation, Silent Spread, that reflects and traces the migratory fast results (within 4 minutes) with 90 pattern of SARS-CoV-2 through 28 individual sculptures. The layout of this piece percent accuracy. was inspired by an SIR diagram of COVID-19 and the decision to present it in Known as RAPID, which stands monochrome was based on NIH/NIAID/RML scanning electron microscopy for real-time accurate portable images of the virus. impedimetric detection, the diagnostic test is built around electrochemical Would you like your photo featured in Image of the Month? impedance spectroscopy (EIS), which Send it to [email protected] transforms the binding event between viral spike protein and human ACE2 receptor into an electrical signal that can be read by a smartphone. The test can be applied to both QUOTE OF THE MONTH saliva and nasal samples, and is able to detect SARS-CoV-2 at low concentrations (1.16 PFU/ml). Perhaps just as importantly, it is also cheap and “The success of LP-GC is an excellent example of a cooperation easy to produce; the electrodes can be between three parties: academics who develop theoretical concepts, mass-produced using screen-printers, hardware developers that create the tools to put these ideas into meaning the entire test costs just $4.67. practice, and users who re-define the workflows in their laboratory High-frequency testing, particularly in to maximally benefit from the new development.” remote and disadvantaged areas, has never looked so likely. Hans-Gerd Janssen, in conversation with Jaap de Zeeuw and Steve Lehotay. Read the full story on page 20. Reference 1. Penn Medicine News (2021). Available at: https://bit.ly/35F7twx c

www.theanalyticalscientist.com 10  Sponsored Feature

A Question of Quality Pharmaceutical analysis – driven by quality control – exists to assure both the safety and efficacy of drug Figure 1. Survey respondents who faced reproducibility challenges shared how often different product groups products. As the world begins to cause issues. emerge from the clutches of the worst pandemic in decades, the role of the pharma industry has never been more visible – nor have its efforts been under so much scrutiny. Here, we present the results of the Pharma Analysis & Quality Control Trends, Challenges and Outlook 2021 Global Survey.

In early 2021 – still very much in the clutches of COVID-19, Merck approached The Analytical Scientist with plans to gather the thoughts of the people “on the ground” in pharmaceutical analysis and quality control. Figure 2. Survey respondents ranked the importance of key trends in pharmaceutical analysis and QC. Their hope? To uncover the major trends and main challenges. “big pharma” companies we know and love Which survey result is most prominent in One survey, over 200 qualified responses, (!) to tiny biotech start ups with a single your mind? and more than a handful of video meetings investigational new drug (IND). What stands out most for me is the later, we are pleased to be able to share the Layered on top of those major trends section on reproducibility and validation final report with you: tas.txp.to/0621PQC in 2020 and 2021 is the COVID-19 (see Figure 1). To whet your appetite, we caught pandemic, which has had its own In terms of reproducibility, there up with one of the minds behind the influence in shaping current trends and are evidently more challenges facing survey – Wayne K. Way, Global Strategy driving new ones. analytical labs than I initially suspected Pharma Analysis & QC, MilliporeSigma With so many trends and challenges – and the issues don’t appear to stem – to explore a few key findings from the pushing and pulling, it felt like a great from one particular tool or application. report in more detail. time to grab our survey camera to take The frequency of reproducibility issues – a snapshot of the industry as viewed by daily and weekly for some respondents – What were the motivations behind those in pharmaceutical analysis and QC. was alarming. the survey? And though we are clearly interested in the In part, I was surprised by the results We were already well aware that the results of the survey, our intent from the because they went against my own pharma value chain has been rapidly evolving start was to create something that would experiences in a pharmaceutical testing over the last 5–10 years – driven by several have value for the industry as a whole – lab – admittedly 20 years ago (see key trends: i) the emergence of biologics very much in line with the aforementioned “Meet Wayne Way”). Although we had from a niche position to a more mainstream trends of partnerships and collaboration. our challenges and out-of-specification market, ii) the rise of partnerships and Indeed, we also sought out an results, I don’t remember them happening collaboration, which essentially represents independent partner with a global that frequently – but maybe I am just a shift in the pharma development mindset, reach – The Analytical Scientist – to help remembering the good parts! Joking aside, and iii) an increasingly diverse spectrum of conduct the survey, analyze the data, and I think the challenges likely reflect the organizations – from the large multinational prepare the report. increasing complexity of today’s drugs and Meet Wayne K. Way formulations – in particular, the increasing surprising at all – especially as we try to proportion of biotherapeutics – which manufacture our way out of a pandemic. inevitably leads to an increasing portfolio of I was surprised by the fact that potential problems. Method transfer from outsourcing ended up so low on the list; development to QC is also likely to be more perhaps it’s because it’s not considered challenging. Finally, time and staff pressure a “trend” as such but perceived as being may be another driver. I was so interested in a steady state. in this section that I reached out to Michael After I received my PhD in analytical Dong, Principal Consultant at MWD You’d already identified partnerships chemistry back in the mid-90s, I joined Consulting, for some perspective: “One of as a global trend; one survey question a small contract testing lab. At that the significant challenges to the separation explored the types of partnerships time, the company didn’t specialize scientist is the efficient development of organizations are engaging in – in pharmaceutical services but that stability-indicating HPLC methods for the any surprises? side of the business was growing, so determination of diastereomers in multi- As you say, we knew partnerships and the company decided to establish a chiral new chemical entities (NCEs).” collaboration were important to pharma, dedicated division – in part because the Whatever the drivers, when I look at those but I was somewhat surprised by the scale industry was so strictly regulated. I led results, I can only imagine the frustration... presented in our snapshot! the team in that new division and we The fact that a third of respondents did all the work you might expect in an We asked respondents to rank important partner with other pharmaceutical industry dominated by small molecules trends affecting pharmaceutical analysis companies is good to see – and we’ve – including stability testing, method and QC – did the results (see Figure 2) seen some very public examples of development, USP testing. And I also meet your expectations? meaningful collaborations in the huge hosted my fair share of FDA audits, so When I look at the data, the top three – effort to vaccinate the world. I’m familiar with that experience… real-time release, the need for higher I was a little surprised to see vendors After some years, I brought my throughput, and continuous and suppliers so high up the pharmaceutical background and manufacturing – are all list. Clearly, everyone has experience to Sigma Aldrich and then related to greater speed Download the relationships with a to MilliporeSigma, Bellefonte, PA, USA, and/or efficiency – Survey Report! number of suppliers – an affiliate of Merck. getting the drug To download your free copy and it is rewarding to see out into the world of report, which also explores that we have evolved faster. And that is not partnerships, productivity from transactional bottlenecks, budgets, and exchanges to fully- more, please visit: fledged partnerships in may be perceived more as a service than tas.txp.to/0621PQC people’s minds. a partnership... The low ranking of contract research and In hindsight, were there any questions contract development and you wished you had included? manufacturing organizations It’s always a balancing act to write a survey (CROs and CDMOs) had us scratching which is short and succinct to allow for easy our heads a little. But when we dug into completion but have the depth to actually the data, we found that CROs/CDMOs realize meaningful results. In hindsight, I were more highly ranked by emerging think we achieved this. If I had it to do biotech companies than by large over again, I would ask more questions pharma, which makes some sense. The around the trends, possibly digging deeper relationship between pharma companies into automation and digitalization. Also, and CROs/CDMOs is complex, so we the issues regarding reproducibility can only make educated guesses as to and validation of products would be the reasons. But, for the bigger players, interesting, since the challenges in those outsourcing a handful of analytical tests areas seem larger than I expected.

sigmaaldrich.com/PharmaQC 12  In My View In My View Experts from across the Living in a world share a single strongly held opinion Material or key idea. World

Material choice is imperative to sample prep success; here are some of the key considerations that chemists must make – and why

By Verónica Pino, Professor of Analytical Chemistry, Chemistry Department and Researcher at the University Institute of Tropical Diseases and Public Health, University of La Laguna, Spain. Committee Member of the EuChemS- DAC Sample Preparation Study Group and Network.

There is no doubt that many of the current advances in sample preparation methods, particularly in the microextraction field, have taken place in parallel with those occurring in material science. As a result, a variety of novel solvents – such as ionic liquids, magnetic ionic liquids, and deep eutectic solvents, among others – have facilitated the development (or improvement) of several liquid-phase (micro)extraction methods. In the same manner, solid materials such as magnetic nanoparticles, carbon-based materials, groups in their structures that facilitate solvents and sorbents for extraction; metal-organic, and covalent-organic further interactions with target analytes, these help us to develop more sustainable frameworks are starting to occupy a and so on. The full list is very impressive analytical methods. And if these reasons privileged position as efficient sorbents indeed – and these materials can also seem insufficient to recognize the in many solid-phase (micro)extraction be combined to form composites, giving impact of materials science in analytical approaches. further mixed or combined properties. chemistry, then consider the interest that The success of sample prep materials Another important aspect to consider such multidisciplinary work can provoke relies on designing and tailoring these when using novel materials in (micro) from other fields! materials to possess certain properties. extraction methods is the resulting But we cannot be blinded by success; Some properties are desirable for increase in the solvents and sorbents it is important not to lose sight of specific analytical applications. They available to us. This expansion is clear what must still be improved. Simply can be tuned to be hydrophobic or and helps us to overcome limitations incorporating a material into an hydrophilic, water-soluble or insoluble, associated with traditional approaches. analytical method does not make it with a modulable degree of porosity and Additional characteristics of “greenness” interesting or novel. The real questions surface area, with certain functional are also associated with many novel we must ask when discussing novel Experts in Reproducibility “We cannot be blinded by success; it is important not to lose sight of • RP-BioLC (U)HPLC what must still be • High Recovery IEX improved.” • Highly Efficient HIC & SEC materials are: does the incorporation of the material improve the performance of the resulting analytical method? Does the material perform better than others (including commercial materials)? Is the proposed material truly green (if claimed) in terms of its preparation and potential cytotoxicity? Does it possess adequate inter-batch reproducibility? And, when using composites, is each individual component necessary? Was the complexity of the final composite really needed? If we do not pay due Discover more at www.ymc.de attention to these concerns, then we run the risk of missing our opportunity to improve the sustainability and performance of our approaches with novel materials. These considerations the case; much research opts to use any Chemical Society (DAC-EuChemS) ensure that chosen materials are fit for available material in seemingly random and includes three working groups purpose and provide us with workable applications, irrespective of its suitability (WG): 1. Science and Fundamentals, results in a way that isn’t detrimental to and design. It is our responsibility to 2. Automation, Innovation and us or our environment. adopt strong criteria in our studies rather Entrepreneurship, 3. Information Then there are also the questions we than succumbing to research trends. Exchange and Networking. must ask beyond the remit of analytical We must not ride the bandwagon – we chemistry. Were all materials properly must guide it! We are visionaries and The Sample Preparation Network synthesized and characterized? Was the our expertise is invaluable. Let us use welcomes new European and non- yield reasonable? Was the preparation it for the betterment of our field – and European regular members. Membership method affordable and sustainable? The our planet. And why not use the benefits is open to individuals who subscribe to list goes on. gained to support other areas of research the objectives of the network and who are Overall, decisions regarding the use – say, material science – while we’re at it? professionally engaged in or associated of novel materials in sample preparation with sample preparation. should be based in their design Sample Preparation Study Group and and selection for specific analytical Network belongs to the Division of For more information please visit: applications. Sadly, this is often not Analytical Chemistry of the European https://www.sampleprep.tuc.gr/en/home

www.theanalyticalscientist.com 14  In My View

such as food products. Enhanced Sample separation is often key to extracting the necessary level of information. “A relationship Preparation in This is true for both food quality and authenticity applications, where even less Multidimensional untargeted approaches are generally applied, and for food safety issues, where investigated is the Analysis targeted and ultra-trace techniques are used. contribution of Multidimensional separation techniques are popular in both of multidimensional these settings (targeted and untargeted analyses). By coupling separation chromatographic techniques, we can enhance peak capacity at each separation step, systems to the providing higher-level information (often multi-level) than single- optimization of dimension approaches. These multidimensional techniques often sample preparation involve chromatography coupled to MS or other informative detectors. steps.” The literature details plenty of fascinating and powerful approaches to increase the separation power of rarely regarded as an extra separation chromatography systems. However, dimension that can contribute to the the impressive development of overall multidimensionality of a system. such analytical systems in terms of In this sense, sample preparation is a sensitivity, selectivity, and separation low-resolution technique that provides power has led people to underestimate a limited additional peak capacity but the role of sample preparation. It may often an essential contribution to the be seen as an ancillary, undesirable, separation power of the process as a and time-consuming step. Many whole. Dedicated sample preparation would rather opt for sophisticated and can therefore simplify the overall Innovative sample prep expensive instrumentation than spend analytical method by reducing our approaches often hide in valuable time optimizing and validating need for sophisticated instruments and the shadows of flashy new the sample preparation step. increasing the power of an analytical techniques – but it’s time for The sample preparation community method. This is particularly useful in them to step into the light is very active in proposing brilliant the context of complex food matrices. I and novel solutions. These include find it wild to think that any researcher By Giorgia Purcar, Professor, Gembloux new sorbent materials and more rapid, would willingly (or perhaps, in some Agro-Bio Tech, University of Liège, miniaturized, and efficient purification cases, unwillingly) pass up such a Passage des Deportes 2, 5030 Gembloux, methods, often with desirable “green” valuable opportunity to enhance their Belgium, and WG1 Leader of the qualities. The potential of these new research capabilities. EuChemS-DAC Sample Preparation approaches is often not fully expressed A relationship even less investigated Study Group and Network due to the lack of analytical techniques is the contribution of multidimensional powerful enough to support their chromatographic systems to the Many analytical problems require high- evaluation and provide evidence to the optimization of sample preparation performance techniques, especially wider community. steps. How? For instance, through when dealing with complex matrices In short, sample preparation is the chromatographic separation of a In My View  15

class of interfering compounds, thus and beyond. In this context, the sample Analytical Chemistry of the European reducing the need for extensive sample preparation community has created, Chemical Society (DAC-EuChemS) preparation steps. A good food-based within the Division of Analytical and includes three working groups example is the problem of mineral oil Chemistry (DAC) in EuChemS, a (WG): 1. Science and Fundamentals, contamination, which requires both Sample Preparation Study Group 2. Automation, Innovation and sophisticated analytical techniques and and Network that aims to promote Entrepreneurship, 3. Information extensive sample preparation. advances in the field and facilitate Exchange and Networking. A closer interaction between communication with other areas of multidimensional separation and analytical sciences. We hope that our The Sample Preparation Network sample preparation could increase the DAC-EuChemS Sample Preparation welcomes new European and non- dimensionality of entire methods in a Study Group and Network will play a European regular members. Membership very dedicated and fit-for-purpose way – fundamental role in building a fruitful is open to individuals who subscribe to allowing us to reach higher and achieve alliance between sample preparation the objectives of the network and who are more ambitious goals. Collaboration and multidimensional techniques. professionally engaged in or associated between these two inherently related Together we will reach for the stars! with sample preparation. communities will increase the amount of information extractable from any Sample Preparation Study Group and For more information please visit: analytical method – in food analysis Network belongs to the Division of https://www.sampleprep.tuc.gr/en/home

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efficiency, flexibility, and speed. In this way, is created. Because there is the ability to Green Food we achieve results of consistent merit, but develop new chemistries that are more with much reduced environmental cost. benign, chemists are obligated to do so.” Analysis: The In sample preparation, we have Therefore, researchers have a responsibility implemented new solvents and integrated to be aware of the valuable links between Missing Link? extraction processes that are more benign in green chemistry and success in food analysis. terms of energy and solvent consumption. My take-home message: we know how we Green analytical chemistry We have also searched for solutions across must proceed and we know what changes we principles could play an omics platforms to help us determine food must make. It is not only our duty, but also important role in helping us hit constituents and nutrients at molecular an invaluable opportunity to give a future key sustainability goals – so levels while protecting the planet. Green to the generations that will come after us. what are we waiting for? chemistry and GAC principles not only promote food security, but also help us strive Sample Preparation Study Group and for many of the individual SDGs, such as Network belongs to the Division of SDG 2 (Zero Hunger), SDG 3 (Good Analytical Chemistry of the European Health and Wellbeing), SDG 6 (Clean Chemical Society (DAC-EuChemS) and Water and Sanitation), SDG 7 (Affordable includes three working groups (WG): 1. and Clean Energy), SDG 8 (Decent Work Science and Fundamentals, 2. Automation, and Economic Growth), SDG 9 (Industry, Innovation and Entrepreneurship, 3. Innovation and Infrastructure), SDG 11 Information Exchange and Networking. By Elena Ibañez, Research Professor in the (Sustainable Cities and Communities), Foodomics Laboratory at the Institute of Food SDG 12 (Responsible Consumption and The Sample Preparation Network welcomes Science Research (CIAL-CSIC), Spain. Production), SDG 13 (Climate Action), new European and non-European regular SDG 14 (Life Below Water), and SDG members. Membership is open to individuals I’d wager that most people have thought 15 (Life on Land) (3). who subscribe to the objectives of the network about the 2030 Agenda for Sustainable Without any doubt, food analysis can and who are professionally engaged in or Development adopted by all United drive renewed approaches in agricultural associated with sample preparation. Nations Member States in 2015, as well development, food processing, food as its 17 Sustainable Development Goals security, nutrition, and health, thus For more information please visit: (SDGs) and how they can impact our promoting the sustainable development https://www.sampleprep.tuc.gr/en/home world and humanity’s future. Six years on, of nations. Moreover, GAC strategies UN reports agree that we are not on track promote not only safer, cheaper, and References to achieve these SDGs by 2030. We can all more sustainable analytical methods for 1. Independent Group of Scientists appointed by the make individual contributions, but strong food analysis, but also more affordable Secretary-General, Global Sustainable Development and cooperative commitment at all levels analytical procedures that can benefit Report 2019: The Future is Now – Science for is needed, including the upper echelons society. Surely we would all benefit from Achieving Sustainable Development, (United of science (1). The question I’d like to ask these strategies in our day-to-day research! Nations, New York, 2019). Last accessed June 15th is: how can we push toward the SDGs in Awareness of green chemistry and 2021. Available at: https://bit.ly/3xqqr5V our everyday scientific work? GAC principles in food analysis has 2. B Gilbert-López et al., “Green foodomics. Towards a Our group at the Foodomics Laboratory increased exponentially in the last 10 cleaner scientific discipline,” TrAC Trends Anal has made attempts to implement the “Green years, with more than 80 percent of the Chem, 96, 31 (2017). DOI: 10.1016/j. Foodomics” framework in our research publications including the terms “green trac.2017.06.013.3. for some time (2). A lot has changed chem*” and “food analys*” since 2010. I 3. D Ballesteros-Vivas et al., “Green food analysis: since we decided to integrate Green hope this awareness expands as we move current trends and perspectives,” Curr Opin Green Analytical Chemistry (GAC) principles forward. As Paul Anastas mentioned Sustain Chem, 100522 (2021). DOI: 10.1016/j. in our laboratory. We have modified our in his inspiring 1999 publication (4): cogsc.2021.100522. analytical methods to fulfill the GAC “With knowledge comes the burden of 4. Paul T. Anastas (1999) Green Chemistry and the requirements, but preserved key qualities responsibility. Chemists do not have the Role of Analytical Methodology Development, for method success, such as accuracy, luxury of ignorance and cannot turn a Critical Reviews in Analytical Chemistry, 29:3, sensitivity, reproducibility, simplicity, cost, blind eye to the effects of the science that 167-175, DOI:10.1080/10408349891199356.

Sponsored Feature  17

disseminate chemical knowledge – and our offerings! For example, a new hub A Collaborative all of this work supports that. We’ve been that offers guidance for interviews and following our charter and empowering preparing CVs, as well as self-assessment Community our members for over 180 years – and tools, will be available to our members for Everyone we’re not stopping now! soon (watch this space). Could you tell us more about the Do you receive any positive feedback on The Royal Society of Chemistry membership categories? your offerings? (RSC) extends a warm helping First is our student membership, which We collect feedback through annual surveys hand to its members throughout is available at a heavily discounted rate to – and the results are overwhelmingly their careers; we spoke with give them the best start possible in their positive. In 2020, 92 percent of responding Jessica Evans to find out more... careers! Then we have our professional members said they were proud to be a categories: associates, members, and member! And 91 percent agreed that we Tell us about yourself and fellows. Associate memberships are advancing excellence in the chemical your role with the RSC are given to those who have sciences. Of course, we also receive some I studied chemistry at Reading just graduated or obtained an constructive criticism – we value this just as then started work analyzing equivalent level of expertise much as the positive feedback as it helps sports supplements. It was through their working life to us to remain relevant and ensures that we exactly what I wanted to date; members have three can continue being a diverse and inclusive be doing and I loved it, but I years of post-study experience; community in the chemical sciences. decided that a more people-facing and fellow-level memberships are Nessa Carson, a member and senior role might be more suited to me. The RSC given to those with at least five years of automation scientist at Syngenta, also was an obvious choice from there, and the experience in a senior role. For people who provided this great feedback about our role of Membership Recruitment Executive don’t yet have a formal qualification or the careers services: “When people ask me why suits me nicely! In addition to being part required years of experience, or who are they should join the RSC, I always tell them of a professional body in the field I’m otherwise involved in the chemical sciences, about the amazing career advice. I think a lot passionate about, I’m also able to support there’s the affiliate category. Members of of people forget that careers support is part other chemists flourish in their careers. I’ll any category benefit from our support of your membership, but it totally is and you have been here for two years in July! irrespective of their working sector. can talk to the team at any time you like.”

What makes the RSC so special? How do you support your members Great feedback! And what else would you I think many people will know us for our through all stages of their careers? say to somebody thinking about joining? publishing work (we have 46 journals Our members work in a fast-paced I’d tell them that we provide support for spanning the breadth of chemical sciences), environment that constantly evolves. We everyone – no matter their career stage, but the RSC is known for a wide range recognize that professional development is sector, or location. And, more than that, of activities. What makes it so special? All key to career progression in this environment; we offer a welcoming community that of these activities are supported by our professional awards offer a framework to encourages conversation and collaboration fantastic member community! And we also ensure that members develop their skills and between talented and like-minded act as an advocate through our policy work. knowledge in a rounded and effective way. individuals. Community is what we’re all In the latter endeavour, we are striving to Chartered Chemist, Chartered Scientist, about – and our members seem to agree! become even more active with bigger Chartered Environmentalist, Chartered After all, 77 percent of them said that they contributions to broad and important goals, Manger and the Professional Registers are felt that they “belonged” with the RSC in such as increased diversity and sustainability. awards that highlight important soft skills our 2020 survey. Our research campaigns – a prime example and facilitate continuous development at To hear more about our members’ being “Breaking the Barriers,” which every career stage. experiences, please visit our website (rsc. focussed on gender equity in academic Then we have our mentoring scheme and li/memberstories) or tune in for one of our publishing – are evidence of those efforts. peer support groups – designed to help our monthly Instagram or Twitter live sessions. Part of our royal charter is to maintain members share experiences and guide one I’m having a blast chatting to the attendees and enhance professional standards and another. But we’re constantly developing and I look forward to seeing you there!

http://rsc.li/membershipcategories 18  Sponsored Feature

years ago; back then, scientists had to bioassays can be used to determine Native LC-MS: build and customize their interfaces to some of the functional characteristics perform fit-for-purpose analyses. We’ve of biomolecules, and surface plasmon From the Lab to the also seen huge amounts of progress resonance can help paint a picture in the development of software and of a molecule’s binding and affinity Manufacturing Floor resources to support such analyses, kinetics, but native LC-MS provides significantly accelerating and simplifying additional insights – and much faster. How is the characterization of data processing. Altogether, native Using this approach, you can quickly non-denatured biomolecules LC-MS has become a much more decipher the high-order structure of a driving scientific progress in approachable technique that even given molecule as well as the covalent industry and academia? scientists with little MS training can use. complexes it forms. And the basic architecture of a molecule can easily The use of native LC-MS has grown In which applications does the be revealed with native LC-MS. significantly in the last decade. But what technique shine? continues to make this tool so useful Rob Haselberg: In a biopharmaceutical What specific challenges does native for professionals in academia and context, native LC-MS helps provide LC-MS present? industry? Heidi Vitrac, an Applications a better understanding of proteins. Vitrac: The hyphenation of LC with MS Scientist at Tosoh Bioscience, and Rob Take monoclonal antibodies (mAbs), is quite different from any other optical Haselberg, an Assistant Professor at the for example; their behavior can be detector. The biggest difference pertains Vrije Universiteit Amsterdam, share analyzed on a molecular level – helping to the need to switch from the liquid their perspectives on the technique, to determine how quickly they bind to phase to the gas phase, which is achieved and explain why it suits the needs of receptors and providing insight into their in the ion source of the MS instrument. the curious minds behind blue skies physicochemical properties. There’s a And that makes optimization of the research, as well as the practicalities of lot of information that can be extracted mobile phase used in the LC separation biopharmaceutical manufacturing. with the native LC-MS approach. a critical component to the method Vitrac: Thanks to various improvements development process. And because What is “native” LC-MS? in sensitivity, resolution, and mass native LC-MS often happens under Heidi Vitrac: Native MS refers to the accuracy of mass spectrometers, native physiological conditions (i.e., neutral pH), process whereby large biomolecules LC-MS is now routinely used in many you cannot use organic solvent or acid in and complexes can be transferred pharmaceutical and biotechnology the mobile phase, which adds to method from a three-dimensional, functional laboratories for the analysis of mAbs development challenges. Notably, native existence in a condensed liquid phase and other protein therapeutics to assess LC-MS is not restricted to size exclusion to the gas phase via the process of purity, and profile antibody glycosylation. chromatography; indeed, several other electrospray ionization (ESI). Native But the technique is not restricted to modes of chromatography can be MS allows solution state non-covalent protein analysis. It can also be applied used in combination with native MS, protein interactions to be maintained in to macromolecular complexes, including including capillary iso-electrofocusing transmission into the gas phase of the drug-macromolecular complexes, lipid- (cIEF), hydrophobic interaction mass spectrometer. When hyphenated protein interactions, and DNA-protein chromatography (HIC), and anion with LC, native LC-MS refers to the interactions. By preserving the native exchange chromatography, as recently analysis of biomolecules in their native, structure of the biomolecules of interest, demonstrated for various mAbs. non-denatured state. native LC-MS allows us to determine the There has been a steady increase mass of intact assemblies, their precise Can native LC-MC be further improved? in the use of native LC-MS over the stoichiometry, direct interactions Haselberg: Sometimes the practical past 10 years, largely driven by the between subunits, and the strength of questions we have are greater than development and expansion of high- inter-subunit interactions. the technology practically allows resolution mass spectrometers from us to explore. So, compromises various vendors. Setting up instruments Can other technologies achieve the between separation and detection and running native MS experiments is same goals? may occasionally be needed. From an not as daunting a task as it was twenty Haselberg: Put simply, no! Of course, academic point of view, flow rate and Sponsored Feature  19

Meet the Experts

Haselberg: As a lead member of an analytical chemistry group, I work on improving hardware. For example, our group often requires columns that can manage low flow rates, so working with a company like Tosoh Bioscience – who truly understands our scientific needs – is great. They also host user meetings, where we can share our data and any Heidi Vitrac, Applications Scientist, Rob Haselberg, Assistant challenges that we have encountered. Tosoh Bioscience. Professor, Faculty of Science, These sessions are always useful as they BioAnalytical Chemistry, share practical solutions with us too. “I obtained my PhD in Physical Chemistry Vrije Universiteit Amsterdam We have had an enduring collaboration at Paris Descartes University and quickly with the company – a testimony to the developed an enthusiasm for cross- “I received a master’s in analytical services it provides! disciplinary studies around analytical chemistry from the Vrije Universiteit biochemistry. In 2008, I moved to the Amsterdam in 2006 before moving What’s next for biomolecular US and joined the University of Texas on to my PhD, which focused on the characterization? Health Science Center at Houston. I characterization of biopharmaceuticals Haselberg: I predict that we’ll see enjoyed taking on challenging projects using a combination of capillary an increased use of affinity columns and contributing to the advancement of electrophoresis and MS. I’ve always that incorporate the power of mass collaborative work. And that’s what led been interested in biomolecular spectrometry in coming years. Why? me to join Tosoh Bioscience in December analysis and, drawing on that Because it will enable us to purify 2020. With Tosoh Bioscience, I contribute passion, I now work in the VU’s and characterize molecules in a single to the development and optimization BioAnalytical Chemistry group, system. I also think we will see more of analytical methods and processes where much of my research is geared multidimensional chromatography, for the separation, identification, and towards understanding native protein including hyphenation, that will characterization of various biomolecules.” characterization. help deepen our understanding of biomolecules. These techniques will help overcome some of the compatibility issues we face today. Vitrac: Like Rob, I am interested in seeing how multidimensional chromatography spectrometer compatibility also have Where does Tosoh Bioscience fit in? can be used in native MS analysis. It to be considered. But working with a Vitrac: We have accumulated a wealth has the potential to create powerful competent vendor can make a huge of experience in the development of pipelines where the quality attributes of difference in how greatly these factors chromatographic methods – and we are any given biomolecule can be assessed impact research. happy to help optimize any separation. during the various stages of downstream Vitrac: From my perspective, the From academic environments to big processing. I am also curious about new biggest needs are related to data pharma, our chromatography experts applications beyond protein therapeutics interpretation. Biomolecules are can support the growing needs of our and complexes. I look forward to seeing complex and teasing apart the customers. As proven specialists in the the next jump from academia to industry, information on each component of the field, we work with biopharma partners especially regarding the analysis of DNA and complex from a series of mass spectra at all scales – from UHPLC analysis to RNA molecules inside complexes. One remains challenging. Data processing purification at the manufacturing scale. thing is cer tain: our team is truly invested software has improved enormously for Any customer with a separation problem in the success of our customers’ work, biotherapeutics, but further development can simply contact us; we will take the so we will continue to find new ways to will help us better understand “out-of- time to listen and to understand the issue work together to take biopharmaceutical the-ordinary” complexes. before finding the best solution. research to the next level.

www.tosohbioscience.com 20  Feature

THE (LONG AND WINDING) ROAD TO LPGC-MS

Jaap de Zeeuw, Hans-Gerd Janssen, and Steve Lehotay recall the origins of low-pressure GC, highlight the hurdles that arose during development, and discuss its relevance today.

PGC, when coupled to MS, is a fast and robust the analytical community. Today, LPGC-MS is accessible to alternative to traditional GC-MS – but the concept all via a commercialized kit. of LPGC is not new; in fact, the low-pressure route Here, three trailblazers on LPGC’s journey – Jaap de Ltowards faster GC has been known to analytical chemists Zeeuw, Hans-Gerd Janssen, and Steve Lehotay – share how since the 1960s. However, several challenges have stood in the technique managed to persevere despite the hurdles and the way of its widespread adoption. In 2000, Jaap de Zeeuw discuss where it might go next. injected (no pun intended) new life into LPGC with a simple solution – a restrictor that maintained positive inlet pressure Disclaimer: Mention of brand or firm name does not constitute for a wide-bore column. an endorsement by the USDA above others of a similar nature Advocates of the technique have had to navigate a long not mentioned. Lehotay’s views in this article are solely his own and winding road fraught with obstacles, including technical and do not represent the views or position of the USDA or any challenges, commercial pressures, and dismissiveness from other entity.

22  Feature

When did you first Hans-Gerd: Well, the general concept has been around for a encounter LPGC? while – Carel Cramers, Piet Leclercq, and Jack Rijks started research into speeding up GC separations at Eindhoven Jaap: I guess I should kick this one off! The idea first occurred University sometime in the 1970s. Cees Schutjes, the first to me in around 1997 when I was thinking about MS and how PhD student in the field, defended his thesis in 1983, which it commonly uses long 0.25 mm columns. This is a logical included a theoretical treatise of the Golay equation. From this choice when you need positive pressure in the injection port equation the increase of the mobile phase diffusion coefficients and the MS is running under vacuum – you need that length immediately followed as one route towards faster GC. When I to have sufficient restriction. But, as I’m always looking for a started as a student in the Cramers’ group in 1986, the thesis challenge, I wanted to see if it was possible to use a wide-bore of Schutjes was standard information we all had to study. We column with MS rather than the typical narrow-bore. also studied vacuum outlet conditions for their higher speed. Based on the Van Deemter equation, I realized that – But it was only around 1998 that we started doing experiments theoretically – at lower pressure a much higher optimal linear in low pressure GC, jointly with Jaap. velocity could be obtained. I checked the literature for vacuum GC but saw that the setup being trialed was often challenging because the vacuum had to be created in the injection system. Instead, I proposed using pliers to restrict the flow on the inlet side of a metal 0.53 mm i.d. capillary. This approach led to a similar separation but an approximately nine times faster run time – and that simple idea was the basis of what we’ve continued to build upon with LPGC-MS to this day!

Steve: Let’s see, my first email to Jaap was on February 25, 2000 – I had come across the title of a presentation that he made in Gifu, Japan, in November 1999, and Aviv Amirav had given me Jaap’s contact info. In the email, I described how I had already made plans for a summer student, Katerina Mastovska, to investigate LPGC-MS with a quadrupole MS instrument. Interestingly, though, I first heard about “subambient pressure GC-MS” in 1989 from Mark Hail of Richard Yost’s group at the University of Florida when we were both graduate students there.

“ BASED ON THE VAN DEEMTER EQUATION, I REALIZED THAT – T H E O R E T I C A L L Y – AT LOWER PRESSURE A M U C H H I G H E R OPTIMAL LINEAR VELOCITY COULD BE OBTAINED.” Meet the LPGC Experts

Jaap de Zeeuw is currently an international specialist in GC at Restek Corporation. With 41 years of experience in GC capillary technology, he has developed many PLOT columns, developed the first bonded wax column, published more than 100 articles in the field of GC, and is the inventor of fast low-pressure GC using restriction at the inlet. Jaap also made the world’s longest fused silica capillary, for which a Guinness World Record was granted. In 2016, he developed a new technique for coating PLOT columns based on SPIN deposition. Jaap has several hobbies, such as gardening, playing music, scouting, and helping out with his wife’s B&B, and he is currently working on a program for teaching creative thinking to technical/analytical people.

Hans-Gerd Janssen has an MSc and Steven J Lehotay is a lead scientist PhD degree in analytical chemistry from with the USDA Agricultural Research Eindhoven University where he studied and Service at the Eastern Regional later worked in Carel Cramers’ group. After Research Center in Wyndmoor, working at Eindhoven University for almost Pennsylvania, USA. Since 1992, he 10 years as assistant and associate professor, has conducted scientific investigations he joined Unilever in 1999. Janssen has and method development research written more than 200 publications involving improvement in the analysis on theory and method development in of pesticides, veterinary drugs, and other chromatography and MS. From 2004 contaminants in food and environmental to 2019, he was a part-time professor at samples. Steven is the co-inventor of the Amsterdam University, next to his position “quick, easy, cheap, effective, rugged, and as senior scientist in analytical chemistry, safe” (QuEChERS) sample preparation focusing on food analysis, at Unilever approach. And he has been awarded with R&D Wageningen. In 2019, he accepted numerous honors, including the AOAC a part-time professorship at Wageningen International Harvey W Wiley Award. University where his research focuses on According to the Stanford c-score recognition-based analytical chemistry. metric, he resides among the top 0.19 Janssen holds positions in the editorial percent of published analytical chemists. advisory boards of several journals and has served in various scientific and organizing committees for international conferences.

www.theanalyticalscientist.com 24  Feature

Did you immediately was clear that the technology would fit into many recognize the benefits of different application areas. this technique? At this point, Varian took ownership of the product and decided to introduce it Steve: I’ve always felt fortunate that I learned chromatography exclusively for the ion trap MS, with from John Dorsey at the University of Florida at the time. Being application in environmental trace a great teacher, Dorsey always started each chromatography analysis of pesticides and PCBs. class with questions about his previous lecture. After spending I think this was a mistake – it weeks on the theory of chromatography with an emphasis took three years before this to optimize separations and peak resolution, Dorsey began “Rapid MS” offering was also class one day by drawing a chromatogram on the chalkboard made commercially available of two peaks with excellent resolution about two minutes because it didn’t gain the apart. He then asked, “What’s wrong with this separation?” expected revenue. Notably, He brushed aside the aspersions that his drawn peaks weren’t the Rapid-MS instrument perfectly Gaussian, and no one in the class saw a problem. He had a slower data acquisition announced, “It’s wasting time!” rate than others, and I think I’ve never forgotten that moment or that concept, but it seems this put a lot of people off. too many others have. I think too many chromatographers and mass spectrometrists forget that we are all analytical chemists. Hans-Gerd: It’s a good The specialist’s mindset fixates on the power of chemical point Jaap makes here. The separations and not enough on practical matters of sample larger instrument and column preparation, throughput, ease, cost, ruggedness, validation, manufacturers are sometimes and – above all – robustness. When I learned about Rapid-MS a bit risk averse, so even though in 2000, I knew immediately that the restriction capillary was Varian eventually recognized the a brilliant idea and a solution for fast GC-MS. Having entered value of LPGC, it took a while to the “real-world” of pesticide residue monitoring in 1992, I develop. Generally, I think the recognized the benefits of LPGC very quickly. I was taught larger companies try to target to always use a guard column in chromatography, and the idea scientists at conferences and to use the guard column also as a restrictor in LPGC was an hope that they will start to elegant solution that I wish I had considered first! use the new technology and spread the word. Jaap: I immediately recognized the value of LPGC-MS, but That works, but is not a some of those around me did not initially. Once I came up with rapid route. my “simple solution,” I proposed the idea to the management of At Eindhoven Varian – the company I was working for at the time. They didn’t University, we always take interest at first, so I wasn’t able to do any experimentation. had a massive interest I then spoke to some of my esteemed colleagues – namely Carel in fast GC. Over the Cramers, Aviv Amirav, and Hans-Gerd Janssen – and Varian years we have helped became more interested with this expert backing. However, many labs convert they decided it was more of an “academic” pursuit best left to their regular GC to a the University of Eindhoven, and this is when Hans-Gerd and faster run. Although I began working together. we were mostly The initial protocol, based on the 0.53 mm capillary following the route of that had to be squeezed on one side using pliers, seemed narrow-bore columns, really promising so we eventually filed for a patent – the it was clear that other initial design used a short 0.10 mm ID restriction (ca. 60 options like very short cm) coupled with a 10 m x 0.53 mm capillary. We worked columns, columns packed with over 20 external groups who all came back with results with very fast particles, or low- showing the speed benefit of the columns. On top of this, it pressure outlet conditions also Feature  25

“ I I M M E D I A T E L Y had their unique advantages. LPGC was always an option that people liked because it required only R E C O G N I Z E D T H E very small modifications to the equipment. VALUE OF LPGC- Sometimes narrow columns are the MS, BUT SOME OF preferred route, sometimes LPGC is better, sometimes both approaches T H O S E A R O U N D work. But I’d say we very much ME DID NOT recognized the benefits of LPGC- INITIALLY.” MS when it came along.

W h a t additional Steve: My colleagues and I had issues with the very narrow developments restriction capillaries in the Rapid-MS product. We ended up led to the simply connecting two commercially available columns from technique as any vendor (5m, 0.18 mm i.d. guard/restrictor capillary with it is today? a 15 m, 0.53 mm i.d., 1 µm film thickness analytical column), which provided both more robustness and theoretical plates. I Jaap: In the first three years, reached out to all the GC vendors for 20 years about LPGC, there was a lot of noise in and in 2021, Restek finally commercialized a product using the market. Lots of people our column dimensions. saw its potential and wanted Aside from this, there have been many technological to experiment with it – one such advances in the past two decades to continually improve person in particular was Steve. upon the performance and features of LPGC-MS. Most During this period, I recognized notably, commercial triple quadrupole MS/MS instruments some limitations in terms of the were introduced, which provided greater targeted analyte restriction lifetime/maintenance detectability (both sensitivity and selectivity) and faster and the coupling, but never data acquisition speeds. High-resolution MS instruments got the chance to work on have also been introduced, for which LPGC is compatible. that at Varian. However, Improvements of QuEChERS and analyte protectants when I joined Restek streamlined sample preparation and improved peak shapes in in 2008, I finally had GC. The development of a light and reliable capillary column the opportunity to try union also helped make LPGC more practical for shipping out different ways of and installation. making the restriction. I came up with another simple Who should consider solution based on LPGC-MS? making the coupling with PressFit and Steve: I think everyone that is using GC for analysis should positioning this inside consider LPGC-MS. My lab has used it routinely for nearly the injector body. This 20 years now. Why? Firstly, megabore columns are preferable meant the coupling and in routine monitoring using GC because of their much greater restriction were always sample loadability and robustness. LPGC is therefore very at high temperature and useful for rapid, sensitive, and robust analysis of pesticides, in an inert atmosphere. A environmental contaminants, and pretty much any GC- publication was written and amenable analyte that isn’t too volatile. Secondly, LPGC as patent filed, but it never made a product can be used as is in general applications, but, as a it to a commercial product. technique, it also possesses more parameters for investigation

www.theanalyticalscientist.com 26  Feature

and innovation than standard GC-MS. For example, Amirav, “ A DRAWBACK OF Fialkov, and I recently published a paper (1) using resistive LPGC IS THAT heating in LPGC-MS with a short 0.25 mm i.d. capillary YOU NEED TO column that achieved multiresidue analysis in <1 min. My long-term research plans are to implement such methods to BUY A SPECIAL enable ultra-fast monitoring without having to ship samples to “ THING;” T H E a lab, for example. In the short term, one of my projects is to R E S T R I C T O R . A systematically evaluate LPGC-MS in food safety applications using different column dimensions. BIG STEP FORWARD WOULD BE LPGC Jaap: What we basically do with LPGC is trade efficiency for WITHOUT THE NEED speed, but using a robust solution. As long as components that elute at the same retention time can be separated by MS, LPGC FOR A RESTRICTOR is useful. Of course, if isobaric components elute together it will AT ALL” not work. In that case, we need separation by chromatography using highly selective stationary phases, or use other means, such as software tools, to meet the analytical need. It’s also worth noting that there are other ways to speed up MS separations, like working at higher flow, extreme fast programming, and using short, smaller diameter capillary columns. But these do not provide the robustness and loadability of the vacuum GC solution.

Hans-Gerd: LPGC is not unique – it is one of approximately 10 routes towards faster GC. But it is a rather simple technique. In generic terms, I would think it is the preferred method if you have simple separations not requiring a high peak capacity and are using MS detection, while sample preparation is not too much of an issue. In such situations, LPGC can increase your sample throughput up to five or ten-fold.

So why haven’t more people adopted LPGC?

Hans-Gerd: It’s a good question, and we have to be honest here. We clearly thought that all labs would move to faster GC, but nowadays we see that the vast majority of them are still using the classical columns with run times between 30 and 60 minutes. I see two reasons for that. First, I think the need for very fast separation is limited. With a regular run time of say 45 minutes you can easily do 30 samples per day. Many labs will not have that many samples. Feature  27

And we should also not forget that the sample preparation, way around. Grants, patents, and citations usually drive their data interpretation, and paperwork that comes with 30 samples choices. Company scientists are similar in that they spend an per day can be significant. The GC run itself is usually not the inordinate amount of their time on niche applications that are rate-limiting step, and for many the benefits of faster GC are difficult for those customers who complain the most loudly. In simply not worth the investment and risk. my view, they should put more focus on improving efficiency A second reason for the limited acceptance of fast GC is the and performance of their most profitable applications that strong overpromise in literature and by some manufacturers. are taken for granted. Moreover, the scientific publishers and With fast GC there is always a price to pay. Narrow-bore media tend to highlight “sexy” topics, which are promoted by columns offer you a faster analysis speed at the expense of those scientists on their editorial boards. a slightly reduced system reliability. LPGC only works for I can give several other reasons more people haven’t adopted simple separations. Fast temperature programming reduces LPGC. There have been informed criticisms over the years, your separation power a lot. These limitations have not always such as the need for more separation efficiency, analyses of been communicated honestly. volatiles, or problems with column bleed. But one of my biggest frustrations is to notice dismissiveness, disbelief, and Jaap: As I’ve mentioned, some were slow to recognize the disinformation from those who have never studied or tried value of this technology and I think this has been the biggest LPGC themselves. limiting step to progress. Even when people (or instrument companies) did recognize the benefits of LPGC, they couldn’t see the value in pursuing it commercially. In particular, Varian What would you like to see didn’t fully recognize the impact of LPGC having low data for the future of LPGC? acquisition rates. Jaap: In theory, the technique could allow even faster Steve: Right. Varian made the initial mistake of treating separations depending on the temperature programming speed Rapid-MS more as an “introduction device” to their ion trap of the instrument. The 15 m column chosen here could also MS detector, which was not ideal for LPGC due to its slow be replaced for a shorter column – like a 7 m x 0.32 mm or (250 ms) data acquisition rate. Secondly, the product was even a 3 m x 0.25 mm column – as long as vacuum conditions introduced before it was fully studied and optimized, leading inside the separation capillary are created. However, there will to narrow marketing and less-than-ideal column dimensions. of course be other challenges for using such short capillaries, Another crucial mistake was the over-pricing of the product. including sample introduction and focusing. Justifiably, the primary goal of a company is to make money – meeting customer needs is only pursued if it serves this Steve: Do you know what may be even better than LPGC-MS? primary goal – but if more customers had demanded LPGC, Fast GC-MS using supersonic molecular beams (SMB) – also then vendors would have taken more notice. Even so, the 20- known as “Cold-EI.” In SMB-MS, the column outlet is not year time frame of Jaap’s US patent held by Varian and then under vacuum, thus LPGC is not possible using that detector. Agilent certainly put a dampener on commercialization by However, column flow rate can be increased to 32 mL/min, others until now. for example, to provide rapid, high-quality analyses. I would Hans-Gerd also makes good points. Furthermore, LPGC urge anyone in this field to view the application notes and does not work for MS techniques that do not operate with publications from Aviv Amirav. His long and winding road the ion source under vacuum conditions. Also, many volatiles has also been fraught with multiple bad timings (company are already analyzed quickly in standard GC, thus LPGC is consolidations) and human foibles. not going to provide as much gain compared to analyses that currently take 15-60 min. Otherwise, LPGC-MS trades a Hans-Gerd: A drawback of LPGC is that you need to buy a small degree of separation efficiency for speed, sensitivity, special “thing;” the restrictor. A big step forward would be and robustness. LPGC without the need for a restrictor at all. This would Misaligned incentivization is another problem. For example, require the gas inlet system of the GC to be able to work with nearly all academicians choose to study what rewards and/ sub-ambient pressures while still avoiding the ingress of air via or interests them the most. The “novelty” of the idea and the split exit – but it should be technically feasible. technique usually motivates them, and they tend to seek Also with regards to the future, Steve’s work on faster integration applications to suit their preferred tool, not the other methods (summation integration) should be mentioned. The

www.theanalyticalscientist.com 28  Feature

“ IT SHOULD BE Jaap: I couldn’t agree more. Essentially, LPGC-MS can speed OF INTEREST TO up a lot of conventional MS applications where analysis time ANYONE USING MS, is important. It should be of interest to anyone using MS, in IN MY OPINION, BUT my opinion, but the best way to get the word out there is to show the data and let experienced chromatographers speak up. THE BEST WAY TO GET THE WORD OUT Steve: As hundreds of people involved in GC analysis can THERE IS TO SHOW attest, I have discussed LPGC in nearly every encounter with them for more than 20 years! My inclusion of a few slides THE DATA AND about LPGC has been a staple in most of my presentations, LET EXPERIENCED to the point that some people are sick of hearing about it C H R O M A T O G R A P H E R S (and I am definitely sick of talking about it!). I’ve emailed many gurus of GC over the years about LPGC, and SPEA K U P.” when I noticed research or review articles in which LPGC should have been mentioned, I sometimes emailed key publications about LPGC to the GC run time might be relevant for the total duration of your authors to inform them of their oversight. analysis, but in terms of costs it is not the main contributor. As a US federal civil servant, I have LPGC combined with simpler, fully automated, and more reliable no business or financial relationships methods for peak integration is an ideal combination. with anyone about my work, and my motivation is to help others improve Steve: Thanks for mentioning the summation function their chemical analyses and lab integration. Indeed, you are right to point out some of the operations. I admire Jaap de Zeeuw drawbacks of LPGC, so I want to summarize how we’ve been for inventing the restrictor approach, overcoming the current limitations of this technique: i) we have and I’ve wished for 20 years that done high-throughput and easy sample prep with QuEChERS my lab could simply purchase pre- (and now QuEChERSER) since 2003; ii) we also have been connected LPGC columns with the using analyte protectants since 2003 to improve peak shapes and dimensions of our choosing. Now separations for somewhat polar analytes; iii) for the past decade, that this option is available, there is we’ve been increasing selectivity of detection by using MS/MS one less excuse for those who haven’t (for targeted analytes only); and iv) since about 2015, we’ve used tried LPGC-MS. summation integration (for targeted analytes only, too). You also mentioned the “thing” needed for LPGC – that Hans-Gerd: This has been a trying “thing” is merely an appropriate guard column and union, journey, that’s for sure! But we’ve all which are not unusual items. I suppose ferrules for the come away from it having grown and megabore column is another “thing,” but standard columns, developed as analytical scientists along liners, septa, ferrules, nuts, and so on are also “things” that the way. For us, the success of LPGC by the same logic should preclude anybody from doing any is an excellent example of a cooperation analyses at all! In any case, Restek now sells that “thing” in between three parties: academics who develop the same way as any other item, and LPGC has always been theoretical concepts, hardware developers that available as a custom item. create the tools to put these ideas into practice, and users who re-define the workflows in their laboratory Hans-Gerd: Excellent remarks Steve! And it’s this type of to maximally benefit from the new development. Many discussion that highlights, I believe, our passion for this often successful new methods are the result of such three-party overlooked technique. I know only a few users of LPGC, but I interactions. am not aware of anyone who tried it and gave up. The technique works and is reliable and there are certainly more people who Reference could benefit from it. 1. A Fialkov et al., J Chromatogr A, 1612 (2020). DOI: 10.1016/j. chroma.2019.460691

Sponsored Feature  29

profiles, we can detect and identify even comprising replicate data). These methods A Herbal very low-level contamination or sample highlighted quantitative changes in aroma profile constituents. Depending on components and identified compounds not (Fraud) Remedy compound type, these differences could be typically associated with native oregano, in the parts per billion (ppb) level. thereby permitting discrimination between Deploying the Thermo The simple sample preparation native and adulterated oregano. Scientific Orbitrap Exploris associated with HS-SPME is a critical factor GC MS System in the fight in non-targeted analysis, as each successive Time to explore? against adulterated oregano manipulation can alter the sample With the Orbitrap Exploris GC 240 system, composition (2), thereby confounding food manufacturers can assure the quality By Giulia Riccardino, Senior Applications analysis. Finally, the GC-HRMS full-scan data of supplies with unmatched efficiency, Specialist, Thermo Fisher Scientific acquisition capability supports rigor, and confidence. Orbitrap targeted, non-targeted, and Exploris GC systems Adulteration of oregano can be accidental or retrospective data analysis permits rapid change- intentional – the latter being driven by price (1), underscoring its over from EI (for and demand. Leaves from other plants (for broad applicability. spectral library search) example, olive, thyme, marjoram, sumac, to softer ionization myrtle, and hazelnut) are frequently used Proof of the pudding (for example, PCI, for as adulterants because they are difficult to We assessed the confirming molecular detect via visual inspection. Whatever the power of the GC- ions through the mode of adulteration, the fight remains the HRMS approach by use of adduct same: to ensure products are authentic and analyzing the aroma information). The safe. Here, we take a look at the capability profiles of native system also has the of the Thermo ScientificTM OrbitrapTM oregano alongside capability to perform ExplorisTM GC 240 mass spectrometer to oregano samples that accurate MS/MS solve this age-old problem. had been adulterated experiments (for structural to simulate fraud. Using elucidation). Quality of herbs, Quality odors the Orbitrap Exploris GC 240 such as oregano, can be assured by Oregano has a diverse composition of system, we compared full-scan total analyzing the aroma profile with principal phytosterols, pigments, and essential ion chromatograms (FS TICs) for native components analysis and differential oils – complexity that can make analysis oregano, native thyme, and oregano analysis. Finally, Orbitrap Exploris GC comes challenging. But by applying the powerful adulterated with a thyme constituent. with the FreeStyle elemental composition combination of GC and high-resolution Our method clearly discriminated calculator and Thermo Scientific™ Mass accurate MS (GC-HRMS), analysts can between native and adulterated samples: Frontier™ Spectral Interpretation software explore the volatile organic compounds differences related to the prevalence of i) for predictive fragmentation and structural (VOCs) – a new window through which odor components common to both thyme elucidation. All together, these features to assess purity. Indeed, the approach and oregano (for example, thymol, which enable compounds to be identified with provides full-scan data acquisition together is present in the aroma of both oregano unprecedented ease and confidence. with high sensitivity, high resolution and thyme, but at different abundances); (240,000 FWHM), and accurate mass and ii) odor components specific to either References determination (<1 ppm). oregano or thyme. 1. M Diaz-Maroto, et al., “Headspace solid-phase Furthermore, GC-HRMS is compatible The peak differences could be visually microextraction analysis of volatile components of with the headspace solid-phase identified on the TIC readout, but were also species”, Chromatographia, 55, 729-735 (2002). microextraction (HS-SPME) technique analyzed statistically. Statistical analysis tools DOI: https://doi.org/10.1007/BF02491788 (1), which permits a simple, one-step included Thermo ScientificTM Compound 2. D Cavanna, et al., “The scientific challenges in extraction and concentration of oregano’s DiscovererTM software (used for PCA moving from targeted to non-targeted mass volatile organic compounds (VOCs) – analysis, which highlights variation between spectrometric methods for food fraud analysis”, mainly monoterpenes and sesquiterpenes. sample groups) and volcano plots (“V-plots,” Trends Food Sci Technol, 80, 223-241 (2018). By comparing test and reference sample used for identifying changes in large datasets DOI: 10.1016/j.tifs.2018.08.007

www.thermofisher.com 30  Sponsored Feature

a gas cylinder might run out in the their product? Finally, we also consider Nitrogen – middle of an analysis. Sometimes we’d the actual size of the unit; those who need to order a replacement cylinder work in crowded laboratories are likely Whenever, Wherever by special shipment to get it on time, to appreciate the small footprint of the adding unbudgeted costs. Furthermore, Solaris generator. Gas generation presents a safer, interruptions in the gas supply can more cost-effective, and more damage analytical instrumentation. In And what do the Solaris XE reliable alternative to traditional other words, the instrument downtime generators feed? tanks – is it time you made associated with gas interruption is not Our facility relies on two Solaris nitrogen the switch? only inconvenient, but also decreases generators hooked up to in-house air operational efficiency. And that’s why lines. One generator is set up with a Most labs within the analytical field rely on we started looking for gas generators. Waters Acquity UPLC-MS system – the a constant and/or reliable supply of gas. We chose PEAK Scientific’s Solaris XE other with a Thermo CoronaTM VeoTM Traditionally, these gases are transported lab nitrogen generator for a number of RS charged aerosol detector (CAD). and stored in high-pressure cylinders – reasons. Firstly, it is highly compatible “gas tanks” – that increasingly feel like with our existing instruments, such as How has switching to gas generation yesterday’s technology; they are unwieldy, the Waters equipment range. Secondly, specifically helped your work? inefficient, and fraught with issues that it is competitively priced – always a big When we relied on traditional gas modern manufacturers should not need benefit to any lab. Thirdly, PEAK Scientific tanks, we found that our activities could to accept. In-house gas generators offer personnel are highly responsive. That be constrained by cylinder stocks or a safer and cost-effective alternative to latter point was extremely impor tant to gas suppliers. And that’s because most bulk gas supply, but how do you know me when I was deciding which supplier gas supply companies have scheduled if they are the right choice for your lab? to go for – if people can’t communicate deliveries; if we ran out of gas before Here, Kelly Abernethy, Associate in a timely manner when they are trying delivery was due, we had to coordinate an Director, Research and Development, to sell you something, how slow will emergency delivery – more complicated Exela Pharma Services LLC, North Carolina, they be once you have actually bought than it sounds. For example, we had to US, describes her experiences with PEAK Scientific’s nitrogen generation system.

Tell us a little about yourself and what you do... I have been working in R&D at Exela Pharma Services for 12 years. Exela is a specialty pharmaceutical company that focuses on developing, manufacturing and marketing generic and proprietary sterile injectable products. As a pharmaceutical company, quality control is of paramount importance – and we conduct potency and impurity profiling for all of our products. Our analytical science capabilities, including mass spectrometry, are vital to the business.

How do you source gas supplies for your analytical science applications? Until around a month ago, we were using standard gas tanks. And, to be frank, we were struggling; for example, Figure 1: The Solaris XE nitrogen generator Sponsored Feature  31

The Solaris XE Nitrogen Generator

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www.peakscientific.com 32  Feature

EMPOWERING the ANALYTICAL S C I E N T I S T i n EVERYONE

Could the general public prove to be key in the next great phase of analytical chemistry? Say hello to citizen science!

By Michel Nielen, FoodSmartphone Coordinator and Principal Scientist of Wageningen Food Safety Research (WFSR) and Professor of Analytical Chemistry at Wageningen University, the Netherlands

34  Feature

itizen science represents a next logical step in the democratization of analytical chemistry. A central aspect of this endeavor is bringing the lab to the sample – or, put another way, taking measurements outside of the traditional lab environment. CAt the forefront of this effort: designing simplified medical and analytical chemistry tools and making them accessible to citizens. Of course, citizen science isn’t a new concept. A classic example is the pregnancy test, which was developed and made available to the public around 50 years ago. And today’s supermarket COVID-19 tests represent a similar development in response to the current pandemic. At our university, citizens have even supported a project studying the impact of climate change-driven disease migration on local insect populations by catching mosquito samples for analysis by experts in the lab. All this stuff is great – but I can’t help thinking there is so much more we can achieve with citizen science. In the case of COVID-19, for example, these tests should not provide a simple yes or no answer – they should communicate valuable data to stakeholders, such as those in government or industry. And, with modern smartphones, we could use location tracking and timestamping to add even more value with real-time geotemporal maps of the acquired data. Imagine the power of such a tool in the fight against the pandemic? Beyond COVID-19, we could also employ a citizen science approach to fight other plagues of humanity, such as environmental pollution and food fraud. The possibilities are tantalizing and we’re making progress, but where are we headed? How far have we gotten already? And what role does analytical chemistry play in this arena? THE (ANALYTICAL) STATE OF AFFAIRS

Right now, many of analytical chemistry’s contributions to citizen science exist in very simple forms. An example: pH testing of local surface waters – what could be simpler, right? But so many more tools are currently in development to form a bridge between analytical techniques and the public. Our FoodSmartphone project is a great example of a success story on this front (see Box 1: The Food Analyzer in Your Phone). This approach was simplified to the degree that even a teenager can perform a test without training. In fact, we are very close to real-world implementation of citizen science devices in the area of food quality and safety. My colleague Yannick Weesepoel has done some cutting-edge Feature  35

THE FOOD one of our PhD students studied ANALYZER IN aptamers as alternatives to antibodies Y O U R P H O N E on this front, and others s studied DNA-directed immobilization and Michel Nielen answers our questions antifouling. This all plays a pivotal role about the FoodSmartphone project in the development of high-density, high-capacity biosensors. Then there What’s your mission with the were of course groups focusing on FoodSmartphone project? detection with the smartphone camera To bring the lab to the sample and and smartphones used in combination rationalize the food quality and testing with electrochemical devices. strategies in Europe. Current strategies rely on the classic (and limiting) dogma And is the method looking promising? of sample acquisition in the field – on The prototype 3D-printed surface- farms, at border inspection points, plasmon resonance (a label-free bio- in supermarkets – and subsequent interaction technique) biosensor on transport to labs. It can then take a a smartphone is definitely a game day, a few days, a few weeks, or even changer – and it comes with a total cost a month to get results. Many of these of less than 100 euros. What’s more, samples are chosen at random, so we two different 3D-printed microfluidic can waste lots of time analyzing the 99 sample preparation approaches have also percent of these that are destined to be emerged from our collaboration with the safe and compliant. Clearly, this isn’t group at Linköping. Novel algorithms totally effective! We thought: what if were defined for image recognition by key stakeholders like farmers and food the group in Belfast, too. As for the inspectors, and eventually citizens, overall methodology, we previously could conduct the pre-screening developed a smartphone immuno-assay themselves? And that’s what we’re for a biomarkers in milk application aiming to facilitate! with the Ozcan group (see the sidebar: Protecting our Environment – and our Sounds great! And who’s involved in Patients!) in 2014. Back then it took the work? around 90 minutes to finally obtain a The FoodSmartphone project was result. Today, our fastest allergen test the result of a collaboration between takes around 30 seconds. seven leading institutes, including Wageningen Food Safety Research, Has the work attracted much Queen’s University in Belfast, the attention? Spanish National Research Council in Absolutely! Of course, we have great Barcelona, the University of Chemistry relationships with the companies that and Technology in Prague, Linköping our institutes are collaborating with, and University in Sweden, CSEM we’ve attracted massive publicity that in Switzerland, and Aquamarijn is still ongoing! The FoodSmartphone Microfiltration in the Netherlands. project has been picked up by radio and Then there were additional partner television, national newspapers and organizations from industry, like all kinds of Internet sources. We were Barilla (Italy) and Zeulab (Spain), and recently approached by a major food academia too. industry to discuss future collaboration The project was divided into five opportunities on the phone project. This different R&D packages. One such is, naturally, very exciting. Watch this package focused on biorecognition; space for updates!

www.theanalyticalscientist.com 36  Feature

P R O T E C T I N G O U R ENVIRONMENT – AND OUR PATIENTS!

A quick case study of citizen science in the fight for a healthy environment and optimal patient care in Lyme Disease

By Aydogan Ozcan, Professor, the University of California, Los Angeles (UCLA), and an HHMI Professor with the Howard Hughes Medical Institute

Identifying and characterizing microscopic objects in water is critical for many applications. Yet, traditional water monitoring technology relies on in-field sample collection with subsequent laboratory analysis by either manually operated microscopy or optics- based flow-cytometry devices, which are expensive and require expert resources. Our group has developed innovative microscopic analysis and make it detection of a panel of target analytes. holographic imaging flow cytometer accessible for both professional and The current recommended approach – (IFC) prototypes for the rapid citizen users alike. “two-tier serology” – has poor sensitivity and automated identification and Citizen science accomplishes multiple for patients in the early stages of characterization of micro-objects in goals at the same time. First, it allows infection, in which antibiotic treatment water. Our IFC device uses a lens-less participants to learn new skills and is most effective. Our method takes image sensor to capture interference knowledge through direct experience. only 15 minutes, is low cost, and can patterns caused by light passing through Second, the potential geographical be conducted with a handheld mobile and around micro-objects in samples. spread of “citizen scientists” can allow reader using a sandwich immunoassay! Our system computes intensity and us to tackle monitoring problems Importantly, we also incorporated phase images for micro-objects in its spread over large areas – and long a synthetic peptide into our assay’s field of view, which are processed to periods of time, too. In these respects, detection panel: modified-C6, composed extract object features like size, color, environmental issues represent a rich of a C6-like epitope linked to a specific morphology, aspect ratio, and thickness. set of problems for citizen scientists to p41 epitope. This component of our assay Our group has successfully applied address. Our work on phytoplankton elevates the diagnostic capability of our this device to label-free monitoring of compositions (which are sensitive to computational multiplexed vertical phytoplankton compositions and the environmental changes and can lead flow assay (xVFA), further increasing identification of potentially harmful to changes in water conditions that the reliability of our results. This xVFA algae in ocean water, as well as pathogen are dangerous for animals and people was validated through a rigorous clinical identification (such as Giardia and alike) is a great example of such an study using samples from New York and Cryptosporidium) in drinking water. We environmental application. Wisconsin. The result: 90.5 percent are currently working with a company We have also explored the development sensitivity and 87 percent specificity, to commercialize this technology in a of a multiplexed, paper-based sensor which is – to our knowledge – the best practical and cost-effective way. Our that harnesses mobile phones and deep diagnostic performance of a point-of- goal is to democratize high-performance learning to diagnose Lyme Disease via care test for Lyme Disease to date. Feature  37

“ Though the devices are designed to b e s i m p l e , the truth is that it takes a lot of work t o b r i n g analytical capabilities t o t h e public.”

work in the PhasmaFOOD EU-project and also collaborates with Jeroen Jansen’s group (see the sidebar: Enabling Citizen Science). For example, by employing handheld, near-infrared scanners to determine the moisture, protein, and fat content of food products through their packaging at supermarkets. These are simple, low-cost devices that can communicate with smartphones and detect issues, such as adulteration or mislabeling. I imagine these handheld devices will have a bright future in such applications – maybe in as little as two years! Though the devices are designed to be simple, the truth is that it takes a lot of work to bring analytical capabilities to the public. We are used to highly complex tools in academic and industrial lab environments, but simple tests with simple targets are needed when asking untrained individuals to play a role in research endeavors. A key part of this simplification is to identify single targets that can provide useful information about topics of interest. An example might be measuring a single metabolite that represents a metabolic pathway of The phytoplankton analyzer from Aydogan's group interest in a disease versus measuring hundreds of related chemicals across the entire metabolome or microbiome.

www.theanalyticalscientist.com 38  Feature

we must face before citizen science hits the mainstream. The required infrastructure is on its way, though! We should all “ The answer be incredibly excited about this. As for uptake of these endeavors by the public at large, we moving shall have to consider the social and psychological challenges as well. Not everyone will be interested in learning about and forward is performing scientific tests. A direct personal benefit could simplification change this, as would be the case with a specific health check, food quality check, or environmental exposure check. And we – not only of should also prepare for communication issues. Test results will inevitably be challenged on social media, as is the case with the devices, scientific information in general. A strong communication but of strategy will help us to inform users and tackle misinformation at its sources. the entire There is hope, though. The younger generation loves to play and fiddle with gadgets – I’d wager there’s a reasonable chance associated that they would also like to try their hand with analytical protocols, devices. People might be even more tempted by the process if it is associated with causes to which individuals have strong too.” connections. Environmentalists, for example, might lead the way by going the extra mile in obtaining pollution samples for some of the large-scale studies in which citizen science will be a powerful weapon. On this example of pollutants, THE OBSTACLES I would guess that it will take around four to five years for AHEAD citizen contributions to make an appearance. For some other applications like the handheld near infrared We’re seeing the literature booming with portable devices and scanners, however, it might take as little as two years. Along sensors, but – more often than not – when you delve a little deeper the way we’ll need to traverse the dreaded development “valley you discover that there’s, for example, a required incubation step of death” (in which significant financial input is needed to of 60 to 90 minutes, the need for a sample preparation step reach the other side). This will mean validating the methods that uses a high-speed centrifuge, or the use of very expensive in labs with experts, building field trials, producing devices materials. These are clear obstacles to adoption by true citizens. at scale, and so on. Getting these instruments to members of Long time windows can dissuade many from participation, and the public will definitely take some yardage, but we are sure who has a spare centrifuge lying around the house? to reap the rewards when we do. The answer moving forward is simplification – not only of the I for one look forward to the citizen-driven future of science! devices, but of the entire associated protocols, too. We need these approaches to be foolproof, featuring quality-by-design principles that deal with the issues of false positive and false negative results. One effective way to achieve data reliability is to provide some sort of indication of the quality of the results to the user. When results are strong enough to use, these should be automatically communicated to databases or the people responsible for processing them. The inevitable risk of sampling errors will be at least partly compensated by the high numbers of data that become available. A BRIGHTER FUTURE?

We aren’t quite where we would like to be yet. We have some great concepts, but there are problems and bottlenecks that Feature  39

handheld NIR to analyze the health and sustainability of chicken fillets through packaging in supermarkets. We have also developed breath analysis equipment to measure citizen health in any location with the Enose company. This tool could allow us to conduct such analyses outside hospitals, where most people are presumably quite healthy – but this may not always be the case! Enabling citizens to conduct their own measurements is of the utmost importance. In the 21st century, citizens are learning to gather and use their data in innovative ways. Whether it be maintaining health, improving their living environment, or examining exactly what their family is eating, citizens want to know more than ever before. The latest technological developments mean that citizens no longer have to rely on institutions for such measurements. The main challenge: most citizens are not analytical chemists… To be truly powerful, we need measurement technologies that can be used with relative ease. More importantly, these technologies need to be developed in such a way that everyone can trust the result, regardless of their level of expertise. These data may then contribute to a much broader repository of “global data” that empowers scientists. Technologically, citizen science has great potential. Any analytical chemist, ENABLING several universities in the east of the however, knows that every measurement CITIZEN Netherlands. Our ambition is to enable has its limitations. Citizens who haven’t SCIENCE technology providers to reach a much- received laboratory training may not anticipated citizen science measurement understand these limitations. Turning Jeroen Jansen, Associate Professor, the solution alongside academic specialists. all of these non-specialists into “Homo Analytical Chemistry group at the In that sense, the role of the citizen is metans” (measuring humans!) is a big Institute for Molecules and Materials, – thus far – of limited importance. We challenge. But this isn’t just a social Radboud University, The Netherlands, want to change that. science challenge. Having anybody discusses his forays in the world of citizen As part of CSII, we worked with contribute to a large data repository with science Nutricontrol to apply handheld near- centralized chemometrics will require infrared (NIR) sensors, which, for a large effort in calibration transfer, We lead the Citizen Science Innovation example, allow farmers to measure the data matching from different devices, Initiative (CSII) – a joint effort quality of their animal feed in a reliable and data preprocessing to remove non- between analytical departments of fashion. In another project, we used chemical perturbations.

www.theanalyticalscientist.com 40  Sponsored Feature

allowing nested acquisitions and high peak Why was the Cyclic IMS-MS developed? Not To Be capacities. The upshot? Improved data In short, it’s the natural result of Waters’ quality and enhanced confidence in results. constant drive to improve performance Missed: IMS-MS Furthermore, as IMS sits after sample characteristics and functionality of our ionization, the method is particularly valuable analytical technologies. The Cyclic IMS- Kevin Giles explores the for ambient ionization approaches MS system advantages include increased wonderful world of (such as DESI, ASAP or REIMS), IM separation capability, multi-functional IMS-MS, and Waters’ MALDI and MS imaging, where operating modes (providing more in-depth key contributions to it upstream chromatography is molecular characterization) seamlessly not possible. Finally, IMS can integrated in an enhanced Q-Tof MS The power and flexibility of determine gas-phase collision with m/z resolution up to 100,000, mass ion-mobility spectrometry- cross-section (CCS) values accuracy down to 500 ppb, increased mass spectrometry (IMS- of ionized analytes. That’s sensitivity (especially for more labile MS) enables accurate important because CCS values species), and improved dynamic range. determination of a broad range of relate to gas-phase structures and Given all the advantages, I suppose the analytes – from small molecules to large can be used to identify species (through question should be, “Why would you not protein complexes. The workflow and data comparison with theoretically-derived CCS develop such an IMS-MS system?” quality advantages of IMS-MS instruments values) and as an additional confirmation like the SELECT SERIES Cyclic IMS are metric in screening experiments (using And what makes the Cyclic IMS increasingly driving the technology’s use in CCS libraries). For all these reasons, we instrument unique? both high-end research and towards more are seeing increasing interest in IMS-MS. Like Waters’ SYNAPT instruments, routine applications. Kevin Giles, Scientific it has a Q-IM-Tof geometry (all other Fellow, MS Research, Waters Corporation, How have you responded to this manufacturers use an IM-Q-Tof UK, tells us more. increased interest? geometry). Crucially, these instruments Our recent webinar series (see the sidebar feature collision cell placement before Can you summarize the impact of IMS-MS? “IMS-MS Webinar Series”) answers many IM separation, as well as after, which MS is an incredibly powerful technology, common questions and requests for permits separation of fragment ions from which, in combination with IMS, is taken to advice. For example, the “Introduction m/z-selected precursors and measurement the next level. Crucially, the extra power to Ion Mobility and the SELECT SERIES of CCS values to aid structural is compatible with modern analytical Cyclic IMS” presentation outlines the determination. Importantly, the cyclic IMS workflows; IMS gas-phase separation basic principles of ion mobility separation allows multi-pass operation, providing user- timescales sit perfectly between those of and the unique capabilities of the Cyclic selectable resolving power, which we’ve LC separation and time-of-flight (Tof) MS, IMS-MS system. shown to scale from ~65 (for one pass) to >750 (for 100 passes). Such performance is unmatched in a commercial instrument. But perhaps the most unique feature of • Joseph Zaia, Boston University - the Cyclic IMS is its ability to perform IMSn IMS-MS Webinar Series Characterization of glycopeptides acquisitions, analogous to the MSn mode in • Libin Xu, University of ion traps, which allows structural analysis of – View on demand Washington - Lipid Profiling and unparalleled detail. And users get all these MS Imaging features in a remarkably small footprint! • Kevin Giles, Waters Corporation • Ralf Laux, Boehringer Ingelheim - Ion Mobility and the SELECT and Ian Wilson, Imperial College What main challenges were SERIES Cyclic IMS London - MetID and structural encountered during development of • David Ropartz, INRAE - elucidation the Cyclic IMS? Characterization of oligo-/ • Mark Skehel, MRC LMB Cambridge There were a number of challenges in the polysaccharides - HDX/MS Experiments development of the Cyclic IMS device. One critical issue we faced was related to control of ions at the interface between What is the role of the step-wave? Cyclic device between rounds of ion IMS-MS Webinar 1 KG: It efficiently transfers ions from the mobility. Last year, we introduced electron source to the mass analyzer. Note that capture dissociation – a complementary Q&A highlights ions are focused off-axis, then transferred fragmentation technique frequently to the mass analyzer through differential applied to protein and peptide studies; Expert panel: Dale Cooper- apertures. The off-axis design moves ions again, this can be used before or after ion Shepherd and Kevin Giles or thogonally to the main flow of the gas – mobility. Also, we recently implemented and that helps keep the system clean; any surface-induced dissociation (SID), which Can Cyclic IMS both separate structural droplets from the ion source simply pass allows us to dissect the interconnectivity conformers and also isolate/trap them straight through. of non-covalent interactions within native for further investigation? protein complexes. KG: Yes. The unique geometry of this What are the main benefits of IMS? system permits selection of sub- D C-S: When interrogating complex Are multiple pass separation times populations of mobility-separated samples, some analytes (including isomers) compatible with LC? ions, followed by their activation and may co-elute; separation by ion mobility KG: Yes. Typical separations of 2-5 passes reintroduction into the instrument allows us to discriminate between these take less than 100 milliseconds. And even for further mobility separation. We species. Also, IMS supports measurement if you opt for the extreme, say 100 passes, can repeat this sequence in an IMSn of collision cross section (CCS), which can it needs only ~1.5 seconds. experimental design that is analogous be compared with a CCS library, giving to MSn. extra confidence in analyte identification Do prolonged times in the ion mobility – especially for very low-level species. region adversely affect resolution? What is the duty cycle efficiency of KG: No. Although ions spend longer Cyclic IMS? What fragmentation options are in the device, they cannot significantly KG: A crucial feature of our mobility compatible with Cyclic IMS? diffuse radially because they are confined; systems is that, while the instruments are D C-S: The standard fragmentation at the same time though, they are being performing one mobility separation, they method is collision-induced dissociation, separated axially –and this step obviously are also accumulating the set of ions for which we can do either before or after the improves the resolving power of the the next mobility separation. And that’s ion mobility step. In IMSn experiments, system as increasingly longer drift paths, why our duty cycle is close to 100 percent. dissociation can be effected within the hence residence times, are used.

the main ion optical axis of the instrument What research has Cyclic also very much appreciate the raw and the orthogonal Cyclic IMS device. To IMS facilitated? performance power of the instrument. address this, we created an electrode array Some early work we did with David Frequently, this translates into the best in which ion movement could be changed Ropartz at INRAE provides a particularly feedback of all – a decision to purchase a by switching the direction of the traveling interesting example. We used the Cyclic SELECT SERIES Cyclic IMS instrument. waves used for ion propulsion and mobility IMS to separate isomeric reducing People only buy instruments if they separation. This system facilitates ion entry/ pentasaccharides; by combining this believe in their value! exit from the Cyclic IMS, including mobility- approach with higher resolution analysis, selected ejection used in IMSn. IMSn and isotope labeling, we confirmed What is the future of IMS-MS? mobility separation of the anomeric I think there is a very bright future, Are there any unsung heroes in the forms. The application of IMSn to locate both in its growing use as a routine development of the Cyclic IMS? structural modifications in isomeric analytical tool, enhancing workflows Bringing a product to market involves species is potentially very far-reaching! and data quality, and as a powerful many people, and the Cyclic IMS research tool to help unravel some instrument is a testament to the skill and What customer feedback have of the most complex creativity of the entire Waters team in you received? analytical challenges. Wilmslow, UK. But I would like to give All very positive! By participating in Overall, it will play particular mention to Peter Carney, the Cyclic IMS community, we can see a central role in Bharat Chande, John Garside, Paul that analysts’ imaginations have been progressing our McIver, and Peter Nixon, who were pivotal captured by the flexibility of Cyclic IMS customers’ science in developing the first Cyclic IMS prototype operation, both in terms of “dial-up” across a wide range back in 2014. resolution and IMSn capability. They of application areas.

www.waters.com 42 Solutions

Solutions

Real analytical problems Collaborative expertise The Novel applications Return of CannaQAP

Melissa Phillips tells us about NIST’s mission to improve cannabis testing

As the US National Institute for areas. Laboratories participating in Standards and Technology (NIST) rolls CannaQAP benefit from an objective out the second exercise in its cannabis evaluation of their analytical approach quality assurance program (CannaQAP), through comparison of their results to the spotlight is on dried flower – with an other participants and to data provided emphasis on distinguishing hemp from by NIST. “drug-type” cannabis. We caught up with NIST Research Chemist Melissa What response have you had from labs? Phillips to find out more. The response has been overwhelmingly positive! The number of participants What’s the goal of CannaQAP? is exceptional (over 100 labs requested CannaQAP will provide interlaboratory samples for Exercise 1 and 80 percent studies twice a year to the cannabis returned results), and we’ve had lots of testing community (including product feedback thanking us for developing developers, forensics labs, third-party this program and for our commitment testing labs, and regulators). Our QAPs to helping the community improve. are designed to assist laboratories In Exercise 1, which focused on in demonstrating and improving the determination of cannabinoids measurement comparability and/or in hemp oils, almost two-thirds of competence in challenging or emerging our participants were contract testing Solutions 43

www.theanalyticalscientist.com 44 Solutions

are doing all across the USA would be View From the useful to gauge our performance. We signed up for Exercise 1 and found “Laboratories Cannabis Lab it very straightforward – we just processed the samples like any other client sample. participating in Amber Wise, Scientific The preliminary data we have been sent Director at Washington- indicate we were very close to the mean CannaQAP benefit based Medicine Creek and average, which is a good sign. Analytics, tells us about We have now signed up for Exercise from an objective the lab’s experience with 2, and we’re particularly interested in CannaQAP the metals testing aspect. We test for an evaluation of their expanded list of metals but we still don’t We decided to take part in CannaQAP cover all the ones on the NIST list, and analytical approach because we feel it’s important to not many labs test for metals beyond increase the amount of data we have arsenic, lead, mercury, and cadmium. through comparison regarding cannabis in all areas, and I would absolutely encourage we want to contribute to moving the other labs to take part – it’s similar of their results to science forward at every opportunity. to a proficiency test but with fewer Every state that has a legal cannabis consequences. Just process the sample other participants program has their own rules, which also like normal and enter the values in the applies to testing requirements, so we database – easy! There’s really no reason and to data felt having a comparison of what labs not to participate provided by NIST.”

laboratories, and about 10 percent were difficulty for participants compared samples, a second set of marijuana product developers. For Exercise 2, we with Exercise 1, as most laboratories samples (THC > 0.3 percent) will anticipate that we will see a greater will need to extract or isolate the be available to laboratories that can proportion of forensic labs. cannabinoids from the plant demonstrate appropriate DEA licensing materials (not required and/or foreign import permits. These Can you share any for analysis of hemp additional samples will contain THC results from the first oils). As legislation on greater than the amount permitted in exercise? hemp refers to THC hemp, to allow laboratories to evaluate We provided the levels on a “dry- their methods with samples that would preliminary results weight basis,” we normally fail a QC check. to participants in are also including December, allowing an opportunity Why should labs consider signing up? them an opportunity for laboratories to Signing up for CannaQAP allows to review and correct report the measured laboratories to evaluate and improve their any errors. The final moisture in the plant own operations, simultaneously helping report is in preparation, samples for comparison the overall cannabis community increase and we hope to publish it in to NIST and the community. confidence in laboratory measurements. the next couple of months. Toxic elements (arsenic, cadmium, The program operates very similarly to a lead, mercury, and more) that can proficiency testing scheme but without What will the second exercise involve? contaminate cannabis plants based on the judgment and related corrective Participants in Exercise 2 will measure the growing conditions (soil, water, actions of a pass/fail grade. All results cannabinoids, moisture, and/or toxic fertilizers) and processing approach from CannaQAP will be peer-reviewed elements in cannabis plant samples, (grinding) will also be a focus of and made publicly available as published which will introduce another level of Exercise 2. In addition to hemp NIST Internal Reports. The results will Solutions 45

be anonymized so that readers can see products and have the know-how to capabilities, CannaQAP is assisting us the amount of variation between labs anticipate and remedy any challenges in the design and characterization of but not how any specific lab performed in extraction, stability, homogeneity, cannabis reference materials. NIST is (participants will know their own and instrumental analysis that might currently working on the development of performance, of course). An added arise. We are also fortunate to have a hemp plant reference material, which bonus is that remainder samples from colleagues and collaborators such as the we hope to release in early 2022. NIST CannaQAP studies can be retained by National Research Council of Canada is also developing numerous analytical participants and used for quality control that we can reach out to for additional methods based on various technologies in the future, while NIST works to information and troubleshooting if (including LC-MS, GC-MS/MS, develop reference materials. necessary, and a community of cannabis and spectroscopy techniques) that can testing laboratories supporting our be directly transferred to forensic or What makes NIST so well-placed to efforts who are extremely willing to industry laboratories. run such a program? share insights and experience and One group at NIST is working to NIST’s long history of work in the assist us in our mission in any way that develop an objective colorimetric test areas of foods, dietary supplements, they can. that will allow greater confidence in tobacco, and other natural products the quantitation claims of screening has primed us for work on cannabis, What other cannabis-related projects approaches, and another group is despite its unique legal and analytical does NIST have in the pipeline? doing fundamental research toward the challenges. We are experts in analytical In addition to helping labs demonstrate development of an accurate marijuana chemistry as applied to these types of and improve their measurement breathalyzer.

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Quantification of Phthalates in CE & RoHS Compliance Testing

The requirements for a CE mark now Figure 1: Single point calibration and chromatograms (TIC and EICs) for the four phthalates. include the requirements for RoHS compliance, which consists of the is a multi-function thermal sample Phthalate Area RSD MDL disclosure of 4 phthalates: Bis(2- injection system for GC-MS, meeting DIBP 3.2% 21.7ppm Ethylhexyl) phthalate (DEHP), and exceeding the RoHS phthalates DBP 2.3% 21.0ppm Benzyl butyl phthalate (BBP), di-n- testing requirements. butyl phthalate (DBP), Diisobutyl Table 1 shows the RSD (n=8) averaged BBP 4.3% 21.0ppm phthalate (DIBP). A quantification at 3.2 percent, 3 times better than the DEHP 2.9% 14.7ppm method is defined in IEC 62321- 8 by method requirement, with MDLs all a TD (Thermal Desorption)-GC-MS below 25 ppm, 4 times better than the Figure 2: Single point calibration and chromatograms technique. The CDS 6150 Pyroprobe method requirement. (TIC and EICs) for the four phthalates.

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Sitting Down With… Michel Nielen, Principal Scientist of Wageningen Food Safety Research (WFSR) and Professor of Analytical Chemistry at Wageningen University, Wageningen, the Netherlands Sitting Down With  51

Please tell us about your current position. industry at AkzoNobel. That was an In the years following that case, one I’m currently professor of analytical amazing period in my career, because of my PhD students worked on a way chemistry at Wageningen University, I spent a lot of time exploring novel to give a quantitative measurement of and I’m also the principal scientist at the technologies. I managed to convince the selectivity of an LC-MS result. He applied research institute Wageningen them to get one of the first API-MS derived empirical relationships between Food Safety Research (WFSR). As systems, because we were encountering all kinds of molecules in databases principal scientist, I’m responsible for a lot of impurities - thanks to the across the globe and came up with a the strategy of the R&D program – so separation power of the new technique probability function that we could I am involved in things like chairing CZE - and struggled to identify apply and tell the court, essentially, our research committee, coaching talent them. This turned out to be quite an how (un)likely it was that another and advising the board on strategic achievement; we ended up proving molecule could mimic the result given R&D decisions. It’s a multidisciplinary that electrospray ionization could be as evidence. This nicely illustrates what institute, so our activities range from used for not only small molecules and we mean by selectivity. analytical chemistry to toxicology, proteins, but also synthetic polymers. microbiology, and data science. In Later on, we combined this with What keeps you awake at night? addition to these roles, I am also MALDI MS and ended up producing Mainly rather crazy ideas and concepts, coordinator of the European MSCA one of the most widely cited review but in the past court cases kept me ITN project FoodSmartphone and co- papers on the subject. After my time in awake because they were stressful chair of the Recent Advances in Food industry, I came almost full circle in my situations. More generally, I don’t like Analysis (RAFA) symposium series. career by returning to academia. Now, how scientists’ integrity is often called once more, my focus is on selectivity in into question. Your position as an expert What are some of the highlights from analytical chemistry. witness is obviously independent in your career in analytical chemistry? a court case but, quite often, it turns I have a long history in analytical You refer to “selectivity” as a common into a debate about the expert’s own chemistry. I started out four decades thread throughout your career. Can integrity and that of their institution. ago doing my master’s research at you tell us more? It’s something I don’t like to see happen. Leiden University. At the time, we To me, there are three fundamental were developing low-dispersion post- questions in analytical chemistry: What is your key message to analytical column reactors for HPLC – this was What’s in there? How much is in there? scientists nowadays? before LC-MS was invented of course And where exactly is it located? That We need to learn how to improve our – and the idea was to address some of first question is essentially the issue communication with target audiences. the selectivity challenges with current of selectivity and it’s something I’ve People should be aware – whether HPLC-UV methods by performing focused on in many different ways. the audience is a group of scientists, chemical reactions after the column. To illustrate what I mean, I’ll use an marketers, industry people, or the general Alas, these reactors had issues of their example from my time as an expert public – of how to correctly communicate own. We ended up solving a lot of them witness in a specific court case for the the work they are doing. One of the key by developing segmented-flow reactors government. Once, the judge asked things we should be looking to develop that enabled longer reaction times and, how I could be sure that there was no in young scientists is the ability to deal eventually, even performed on-line other molecule on the planet that would with the doubts around lab testing. immunoassay detection after HPLC yield a similar LC-MS signature to the It’s not only an issue in court cases; in for the first time. That was a highlight banned substance under investigation. general society, we see that governments of my early career that has influenced my I’d never thought about it like that. and scientists are being trusted less and later research, combining instrumental Until that point, we’d followed the less. As analytical scientists, we cannot analysis with biorecognition assays. official EU legislation – determining ignore that situation and we certainly After my master’s, I did my PhD retention time, checking against have a role to play in remedying it. From at the Free University of Amsterdam, reference standards, determining the my perspective, it would be beneficial where I became interested in bringing relative abundances of certain ions, and to build aspects such as social science, additional selectivity to HPLC-UV via checking that they didn’t deviate too psychology, and marketing techniques on-line SPE. I then spent 10 years in much. So this really got me thinking. into our analytical education.

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